Cellular stress has been associated with inflammation, yet precise underlying mechanisms remain elusive. In this study, various unrelated stress inducers were employed to screen for sensors linking altered cellular homeostasis and inflammation. We identified the intracellular pattern recognition receptors NOD1/2, which sense bacterial peptidoglycans, as general stress sensors detecting perturbations of cellular homeostasis. NOD1/2 activation upon such perturbations required generation of the endogenous metabolite sphingosine‐1‐phosphate (S1P). Unlike peptidoglycan sensing via the leucine‐rich repeats domain, cytosolic S1P directly bound to the nucleotide binding domains of NOD1/2, triggering NF‐κB activation and inflammatory responses. In sum, we unveiled a hitherto unknown role of NOD1/2 in surveillance of cellular homeostasis through sensing of the cytosolic metabolite S1P. We propose S1P, an endogenous metabolite, as a novel NOD1/2 activator and NOD1/2 as molecular hubs integrating bacterial and metabolic cues. Intracellular pattern recognition receptors NOD1/2, best known as sensor of bacterial peptidoglycans, can also be directly activated by signaling lipid sphingosine‐1‐phosphate accumulating upon perturbed cellular homeostasis, thus linking general stress to inflammatory responses. Altogether, we established that sphingolipid metabolism governs inflammation triggered by cellular stress and identified a central role for S1P in triggering NOD1/2‐dependent inflammation upon stress induction.IL6 and IL8 expression and production correlated with NOD1/2 expression, without considerable differences of cell death in association with NOD1 or NOD2 expression  upon perturbation of cellular homeostasis were compromised in both Nod1/2 dKO and Rip2 KO cells  and NOD1/2 double knockout (dKO). NF‐κB luciferase assays confirmed that perturbation of cellular homeostasis induced NF‐κB activation, while NF‐κB activation and  dKO Fig . Consistlls Figs . AccordiMDMs Fig C and D. et al,et al,et al,ACER1/2, SMPD3,SPHK1 and other key enzymes involved in sphingolipid metabolism  encode the sphingosine kinase  resulted in lower IL8 production upon Shigella infection, suggesting the involvement of S1P in Shigella‐induced NF‐κB activation  were employed for two reasons: (i) Sphingolipid metabolism in cancer cells is dysregulated  or neutral sphingomyelinase (GW4869), uncovering that S1P is mainly produced via the hydrolysis pathway upon ER stress , S1PR2 (JTE013), S1PR1 and S1PR3 (VPC23019), and S1PR4 (CYM50358), as well as the general S1PR inhibitor FTY720 or blocking GPCR signaling by pertussis toxin, were employed. All inhibitors failed to block IL6 production by cytosolic S1P  with the S1P molecule using AutoDock  domain of human NOD1 is critical for the interaction with S1P  or caspase recruitment domain (CARD)‐containing proteins, resembling inflammasomes in vertebrates  and B6N.129S6‐Sphk2tm1Rlp/J (Sphk2−/−) mice were originally obtained from the Jackson Laboratory . These mice were maintained under specific pathogen‐free (SPF) conditions at the Max Planck Institute for Infection Biology in Berlin, Germany. 6‐ to 15‐week‐old female mice were employed for the experiments.B6N.129S6‐Sphk1et al,NLRP3‐GFP  was obtained from Addgene. NOD1‐ or NOD2‐related plasmids were constructed as described , anti‐GFP , anti‐GFP , anti‐RIP2 , anti‐SPHK2 , anti‐phospho‐p38 , anti‐phospho‐JNK , anti‐phospho‐ERK1/2 , anti‐phospho‐p65 , and anti‐NOD1 .l‐glutamine , 10 mM HEPES buffer , pH 7.2‐7.5, and 50 µM 2‐mercaptoethanol . To differentiate THP‐1 into macrophage‐like cells, THP‐1 cells were stimulated with 50 ng/ml phorbol 12‐myristate 13‐acetate  for 24 h and then incubated with fresh medium for another 48 h. HEK293T WT and HEK293T NOD1/2 dKO cells were generated as described  were obtained from tibial and femural bones and generated with DMEM containing 20% L929 cell supernatant, 10% FBS, 5% heat‐inactivated horse serum, 1 mM sodium pyruvate, and 2 mM l‐glutamine. For BMDM stimulation experiments, BMDMs were treated with IFN‐γ (20 ng/ml) overnight before stimulation.The human monocytic cell line THP‐1 was obtained from the American Type Culture Collection  and maintained in RPMI 1640  with 10% (v:v) heat‐inactivated fetal bovine serum , 1 mM sodium pyruvate , 2 mM ueous one solution reagent (Promega) according to the manufacturer’s instructions. Cell viability upon various stimulations was normalized to corresponding controls.Cell viability assays were performed with CellTiter 96 AQShigella flexneri M90T afaE .NOD1/2 KO NF‐κB reporter cells were seeded in 96‐well plates and stimulated with various stressors for 8 h. Then, cells were lysed in lysis buffer (Thermo Fisher Scientific) and luciferase activities were determined with firefly luciferase glow kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. Luciferase activities were normalized to the amount of protein determined with Coomassie Plus kit (Thermo Fisher Scientific). Fold inductions were calculated against normalized luciferase activities of corresponding controls.HEK293T WT and 6 cells were treated with indicated stimuli for 2 h. After washing twice with Dulbecco's phosphate‐buffered saline (DPBS) without calcium and magnesium, cells were collected and suspended in 300 µl of DPBS. Afterward, cells were transferred into 1.5‐ml Eppendorf tubes precooled at −80°C. The lipidomic analysis was performed by Lipotype GmbH. For analysis of sphingosine‐related metabolites, 3 × 106 cells were seeded in Lumox 50 dishes and treated with indicated stimuli for 2 h. After washing twice with 0.9% NaCl preconditioned at 37°C, the membrane of Lumox dishes was placed into tubes precooled at −80°C. 600 µl of quenching solution (dichloromethane/ethanol) was added into each tube. The measurement was performed by Metanomics Health GmbH.For lipidomic profiling, 1 × 102, 0.1% SDS, 1X PhosSTOP, 1X Protease inhibitor cocktail without EDTA). Protein concentration was measured by BCA assay kit (Thermo Fisher) and diluted with delipidized serum to 1 µg/µl. Then, the diluted samples were added into S1P ELISA plates according to the instructions.Quantification of S1P abundances in cell lysates was performed with S1P ELISA kit  according to the manufacturer’s instructions with some modifications. Human neonatal dermal fibroblasts were grown in DMEM full medium. For each condition, 2x T75 flask cells were required. After 4‐h stimulation with various stressors, cells were washed with PBS and collected in lysis buffer  in a Bio‐Plex 200 system according to the manufacturer’s instructions.The mutagenesis was performed with Q5® site‐directed mutagenesis kit  according to the manufacturer's instructions. The primers used in this study were as follows: NOD1 E157A F: 5′‐GCTGCTGGAGgcgATCTACATGG‐3′, NOD1 E157A R: 5′‐AGCTCCTCCTTCTGGGCA‐3′, NOD1 D161A F: 5′‐GATCTACATGgcgACCATCATGGAGCTGG‐3′, NOD1 D161A R: 5′‐TCCTCCAGCAGCAGCTCC‐3′, NOD1 D203A F: 5′‐CATCCTGGGTgcgGCTGGGGTGG‐3′, D203A R: 5′‐AAGATGGTCTCACCCTGC‐3′, L218A F: 5′‐GCTGCAGAGCgcgTGGGCCACGG‐3′, L218A R: 5′‐CGCTGTAGCAGCATGGAC‐3′, R237A F: 5′‐CTTTCGCTGCgcgATGTTCAGCTG‐3′, R237A R: 5′‐TGGAAGAAGAATTTGACC‐3′, H257A F: 5′‐GCTCTTCAAGgcgTACTGCTACCCAGAGCGG‐3′, H257A R: 5′‐AGGTCCTGCAGACACAGC‐3′, E267A F: 5′‐GGACCCCGAGgcgGTGTTTGCCT‐3′, E267A R: 5′‐CGCTCTGGGTAGCAGTAGTG‐3′, H290A F: 5′‐GGACGAGCTGgcgTCGGACTTGGACC‐3′, H290A R: 5′‐AGGCCATCGAAGGTGAAG‐3′, E306A F: 5′‐CTGCCCCTGGgcgCCTGCCCACC‐3′, E306A R: 5′‐GAGCTGTCAGGCACGCGGC‐3′, K324A F: 5′‐GAAGCTGCTCgcgGGGGCTAGCAAG‐3′, K324A R: 5′‐CCACTGAGCAGGTTGGCC‐3′, K328A F: 5′‐GGGGGCTAGCgcgCTGCTCACAG‐3′, K328A R: 5′‐TTGAGCAGCTTCCCACTG‐3′, R340A F: 5′‐CGAGGTCCCGgcgCAGTTCCTGCGGAAG‐3′, R340A R: 5′‐ATGCCTGTGCGGGCTGTG‐3′, R344A F: 5′‐CCAGTTCCTGgcgAAGAAGGTGCTTCTCCGG‐3′, R344A R: 5′‐CGCGGGACCTCGATGCCT‐3′, R350A F: 5′‐GGTGCTTCTCgcgGGCTTCTCCC‐3′, R350A R: 5′‐TTCTTCCGCAGGAACTGG‐3′, D372A F: 5′‐GGCCCTGCAGgcgCGCCTGCTGA‐3′, D372A R: 5′‐CGCTCGGGGAACATCCTC‐3′, R373A F: 5′‐CCTGCAGGACgcgCTGCTGAGCCAGCTG‐3′, R373A R: 5′‐GCCCGCTCGGGGAACATC‐3′, R399A F: 5′‐GATCATCTTCgcgTGCTTCCAGCACTTCC‐3′, R399A R: 5′‐CAGCAGAAGAGGGGCACA‐3′, D423A F: 5′‐GACCCTGACAgcgGTCTTCCTCCTGGTC‐3′, D423A R: 5′‐ATCGTGCAGTCGGGCAGC‐3′, R435A F: 5′‐5′‐CCATCTGAACgcgATGCAGCCCAGC‐3′, R435A R: 5′‐ACCTCAGTGACCAGGAGG‐3′, Q484A F: 5′‐GGAGGAGGTGgcgGCCTCCGGGC‐3′, Q484A R: 5′‐TGGGTGAAGACAAAGAGGCTC‐3′, L495A F: 5′‐AGACATGCAGgcgGGCTTCCTGCGG‐3′, L495A R: 5′‐CTCTCCTGCAGCCCGGAG‐3′, E514A F: 5′‐GCAGTCCTATgcgTTTTTCCACCTCACCCTCCAGG‐3′, E514A R: 5′‐TGGTCACCCCCGGGGCCC‐3′, H517A F: 5′‐TGAGTTTTTCgcgCTCACCCTCCAGGCCTTCTTTAC‐3′, H517A R: 5′‐TAGGACTGCTGGTCACCC‐3′, L540A F: 5′‐CACTCAGGAGgcgCTCAGGTTCTTCCAGG‐3′, L540A R: 5′‐CCCACCCTGTCGTCCAGC‐3′, F544A F: 5′‐GCTCAGGTTCgcgCAGGAGTGGATGCCC‐3′, and F544A R: 5′‐AGCTCCTGAGTGCCCACC‐3′.2+, 1 mM ATP). To elute the proteins, the agarose was mixed with 2 ml elution buffer  for 1 min and the eluted proteins were immediately neutralized with 0.2 ml of 1 M Tris, pH 10.5.HeLa NOD1 or NOD2 cells from 20 T175 flasks were collected and lysed with 5 ml lysis buffer . The lysates were incubated with 2 ml GFP‐Trap agarose at 4°C for 4 h. The agarose was collected with Poly‐Prep columns by gravity flow. Afterward, the agarose was sequentially washed with 50 ml wash buffer I  and 50 ml wash buffer II . To remove heat shock proteins, columns were further washed with 50 ml wash buffer III  and treated with doxycycline overnight to induce NOD1‐GFP or NOD2‐GFP expression. Afterward, cells were stimulated with S1P‐TAMRA (20 µM) in the presence of digitonin (5 µg/ml) for 2 h. Then, cells were fixed with 4% (v:v) paraformaldehyde in PBS, pH 7.4 for 10 min at room temperature (RT). After washing twice with PBS, cells were incubated with 50 mM glycine in PBS, pH 7.4 for 10 min and permeabilized with 0.05% saponin , 1% BSA in PBS for 10 min. Rabbit anti‐GFP antibody (ChromoTek) and Alex Fluor 647‐conjugated goad anti‐rabbit antibody (Thermo Fisher Scientific) were diluted in PBS and incubated for 1 h at room temperature. The super resolution imaging was performed with Nanoimager S (Oxford Nanoimaging). For single particle tracking in live cells, Nanoimager S was pre‐warmed at 37°C. HeLa inducible NOD1‐GFP or NOD2‐GFP cells were stimulated with S1P‐TAMRA (20 µM) in the presence of digitonin (5 µg/ml) for 30 min. Medium was then replaced with fresh DMEM without phenol red. Live imaging was performed with Nanoimager S at acquisition speed of 100fps.GAPDH), and fold changes were calculated against corresponding controls. qRT–PCR data were representative of at least three independent experiments, with at least 2 technical replicates per experiment. The sequences of primers used in this study were as follows: hGAPDH F: 5′‐GGAGCGAGATCCCTCCAAAAT‐3′, hGAPDH R: 5′‐GGCTGTTGTCATACTTCTCATGG‐3′, hIL6 F: 5′‐ACTCACCTCTTCAGAACGAATTG‐3′, hIL6 R: 5′‐CCATCTTTGGAAGGTTCAGGTTG‐3′, hIL8 F: 5′‐TTTTGCCAAGGAGTGCTAAAGA‐3′, hIL8 R: 5′‐AACCCTCTGCACCCAGTTTTC‐3′, mGAPDH F: 5′‐AGGTCGGTGTGAACGGATTTG‐3′, mGAPDH R: 5′‐TGTAGACCATGTAGTTGAGGTCA‐3′, mIL6 F: 5′‐TAGTCCTTCCTACCCCAATTTCC‐3′, mIL‐6 F: 5′‐TTGGTCCTTAGCCACTCCTTC‐3′, hS1PR1 F: 5′‐TTCCACCGACCCATGTACTAT‐3′, hS1PR1 R: 5′‐GCGAGGAGACTGAACACGG‐3′, hS1PR2 F: 5′‐CTAGCCAGTTCTGAAAGC‐3′, hS1PR2 R: 5′‐ACAGAGGATGACGATGAAG‐3′, hS1PR3 F: 5′‐GAGGAGCCCTTTTTCAAC‐3′, hS1PR3 R: 5′‐TCATTTCAAAGGGAAGCG‐3′, hS1PR4 F: 5′‐GACGCTGGGTCTACTATTGCC‐3′, hS1PR4 R: 5′‐CCTCCCGTAGGAACCACTG‐3′, hS1PR5 F: 5′‐AGGAAGCTCAGTTCACAG‐3′, and hS1PR5 R: 5′‐GATTCTCTAGCACGATGAAG‐3′.Total RNA was isolated with TRIzol reagent, as described by the manufacturer (Invitrogen). RNA (1 μg) was used to generate cDNA via the iScript cDNA Synthesis Kit (Bio‐Rad), and qRT–PCR was performed using Power SYBR Green Master Mix (Applied Biosystems) in a StepOne Plus thermocycler (Applied Biosystems). The average threshold cycle (Ct) of quadruplicate reactions was employed for all subsequent calculations using the ΔΔCt method. Gene expression was normalized to glyceraldehyde‐3‐phosphate dehydrogenase . Data represent fold changes (2−ΔΔCT) in transcripts relative to the appropriate internal control (DMSO). Data were generated from 2 technical replicates and at least three independent experiments. TaqMan probes are listed in Table Gene expression was analyzed simultaneously with the 96.96 Dynamic Array Integrated Fluidic Circuits from Fluidigm as previously described  following the manufacturer’s protocol using glycogen as co‐precipitant. Quality control and quantification of total RNA was analyzed using an Agilent 2100 Bioanalyzer (Agilent Technologies) and a NanoDrop 1000 UV–Vis spectrophotometer (Thermo Fisher Scientific). Microarray experiments were performed as single‐color hybridization. Total RNA was amplified and labeled with the Low Input Quick‐Amp Labeling Kit (Agilent Technologies). In brief, mRNA was reverse‐transcribed and amplified using an oligo‐dT‐T7 promoter primer and labeled with cyanine 3‐CTP. After precipitation, purification, and quantification, 0.75 μg labeled cRNA was fragmented and hybridized to custom whole genome human 8 × 60K multipack microarrays (Agilent‐048908) according to the supplier’s protocol (Agilent Technologies). Scanning of microarrays was performed with 3 μm resolution (8x60K) using a G2565CA high‐resolution laser microarray scanner (Agilent Technologies). Microarray image data were processed with the Image Analysis/Feature Extraction software G2567AA v. A.11.5.1.1 (Agilent Technologies) using default settings and the GE1_1105_Oct12 extraction protocol. Expression data were analyzed using R scripts. The data were quantile‐normalized between arrays, and batch effect of purification state was removed by ComBat  overnight at 4°C. Afterward, agaroses were washed twice with PBS and boiled with sample buffer for SDS–PAGE. For lipid‐coated beads IP, lipid‐coated beads were incubated with supernatants for 2 h at room temperature and then washed with lysis buffer for three times and boiled with sample buffer for SDS–PAGE.HeLa NOD1 or NOD2 cells were grown overnight and afterward treated with doxycycline overnight to induce NOD1 or NOD2 expression. Cells of one 10‐cm dish were lysed with 500 µl of lysis buffer  supplemented with protease inhibitors  on ice for 20 min. Lysates were centrifuged at 16,000 × g for 10 min, supernatants were heated with SDS sample buffer at 95°C for 10 min. Proteins were separated on 4–15% SDS gels  and transferred onto nitrocellulose membranes. The membranes were incubated with primary antibodies at 4°C overnight and secondary antibodies at RT for 1 h, respectively. All antibodies were diluted in PBS supplemented with 0.1% Tween‐20  and 5% BSA. Membranes were developed with ECL detection Kit  and exposed with ChemiDoc (Bio‐Rad). The antibody against ACTB/β‐actin was used as loading control.Cells were lysed in RIPA buffer supplemented with protease inhibitor cocktail and PhosSTOP  on ice for 10 min. After centrifugation at 16,000 × Kd values.Binding of NOD1, NOD2, GFP, or NLRP3‐GFP to different ligands was measured by microscale thermophoresis (MST). 20 nM GFP or GFP‐tagged proteins in MST buffer  was incubated with different concentrations of ligands. Immediately, samples were loaded into standard glass capillaries (NanoTemper) and thermophoresis analysis was performed on a NanoTemper Monolith NT.115 instrument  at 22°C. A laser on‐time of 30 s and a laser off‐time of 5 s were used. The experiment was performed in triplicates, and the MST curves were fitted using NT analysis software to obtain the et al,et al,et al,et al,AutoDock Tools  was satisfied. The system was first minimized with the restraints  on the solute (includes the protein and ligand) followed by another round of free minimization. The systems were heated from 0 to 300 K  on the protein and ligand in 100 ps. Then, the unrestrained systems were equilibrated for 1 ns with Langevin thermostat in the NPT (P = 1 atm and T = 300 K) ensemble  with Holm–Sidak post hoc test and ANOVA test of linear mixed model for fix effect of main factors were performed. A two‐tailed P value of < 0.05 was considered to be significant.Statistical analysis was performed with GraphPad Prism v7.03 . Normal distribution has been evaluated with the Shapiro–Wilk test using GraphPad Prism. To determine statistical significance among investigated groups, Student’s GP, AD, and SHEK conceived and designed the study, analyzed the data, and wrote the manuscript. GP designed and performed most of the experiments. JZ, JW, and H‐JM conducted microarray and data analysis. LH performed molecular docking. MD performed molecular dynamic simulation. PM‐A generated knockdown cells. PS participated in the mouse experiments. LL performed Fluidigm gene expression analysis. TAK provided cell lines, plasmids, and conceptual discussions. KE and CA generated HeLa inducible GFP, NOD1‐GFP, and NOD2‐GFP cells. HS performed Shigella infection experiments. NN, YD, MP, and IGB helped with the experiments. All the coauthors commented on the manuscript.The authors declare thet they have no conflict of interest.AppendixClick here for additional data file.Expanded View Figures PDFClick here for additional data file.Source Data for Expanded View and AppendixClick here for additional data file.Review Process FileClick here for additional data file.Source Data for Figure 4Click here for additional data file.Source Data for Figure 5Click here for additional data file.


In complex 1 the Re—Re bond lengths [2.9767 (3)–3.0133 (2) Å] are close to double the covalent Re radii (1.51 Å). The palladium–rhenium carbonyl cluster 2 has not been structurally characterized previously; the Pd—Re bond lengths [2.7582 (2)–2.7796 (2) Å] are about 0.1 Å shorter than the sum of the covalent Pd and Re radii (1.39 + 1.51 = 2.90 Å). One carbene ligand and a diethyl ether molecule are disordered over two positions with occupancy ratios of 0.5:0.5 and 0.625 (15):0.375 (15) in 1. An unidentified solvent is present in compound 2. The given chemical formula and other crystal data do not take into account the unknown solvent molecule(s). The SQUEEZE routine [Spek  and rhenium (Re) led to the isolation and crystallographic characterization of tetrakispalladium(II) hexadecacarbonyltetrarhenium diethyl ether disolvate, [Pd(CSpek 2015. Acta Cr Two other products, triphenylphosphine oxide and the known complex Re2(CO)8(PPh3)2  and rhenium (Re) have important applications in alkane reforming, industrial chemical production, hydrodechlorination and biomass conversion  cation with four coordinated N-heterocyclic carbenes (NHC) lying on a twofold rotoinversion axis, and one [Re4(CO)16] anion. The geometry around the Pd atom is square-planar with one carbene unit being disordered. The C—Pd—C angles range from 86.9 (4) to 97.7 (4)°. The cluster anion lying on the inversion center has a perfectly flat rhombus geometry with the shortest Re—Re bond [2.9767 (3) Å] corresponding to the short diagonal. The other four Re—Re bond lengths [3.001 (2)–3.0132 (2) Å] are also close to double the covalent Re radii –2.7796 (2) Å] are close to the sum of the covalent Pd and Re radii (1.39 + 1.51 = 2.90 Å). In comparison, the Pd—Re bond lengths in the PdRe4(CO)16(μ-SbPh2)2(μ-H)2 cluster –90.135 (6)° and the Pd—Pd bond lengths are 2.9678 (2)–2.99 (2) Å].The displacement ellipsoid plot of 1, each cation is surrounded by six anions and vice versa palladium(II) diiodide 16]2− anion, which is also found in the compound reported here. A search of the CSD found two closely related cluster compounds, viz. bis(tetraethylammonium) hexadecacarbonyl-tetrarhenium hexadecacarbonyltetrarhenium 4  was added to a toluene–acetonitrile mixture  and 1,3-dimethylimidazolium-2-carboxylate . The reaction mixture was refluxed for 1.5 h, then Re2(CO)10  was added, the solution turned dark red and the solvents were removed in vacuo. The solid was washed with benzene (3 × 5 ml) and recrystallized from an acetonitrile–diethylether mixture. X-ray quality crystals of Pd(IMe)4Re4(CO)16·2C4H10O  were grown from a dichloromethane–diethylether mixture at 277 K. 1HNMR : 3.41 , 7.37 . 13C{H} NMR : 36.9 , 123.5 , 168.0 , 197.7 (CO), 198.7 (CO), 201.1 (CO), 218.6 (CO) IR : 3152 , 1998 (vw), 1974 (vw), 1955 (m), 1927 (vw), 1912 (vw), 1881 , 1858 (vw), 1575 (vw), 1465 (w), 1400 (vw), 1332 (vw), 1229 (m), 1131 (vw), 1083 (vw), 1013 (vw), 845 (vw), 736 (s), 701 (vw), 681 (m), 600 (w), 577 (s), 560 (vw), 508 (vw), 496 (vw), 464 (w), 436 (vw), 411 (w).Under a nitrogen atmosphere, Pd(PPh4Re2(PPh3)4(m-CO)8(CO)2 suitable for X-ray diffraction analysis were obtained from a yellow benzene solution, after several days, by slow ether diffusion into a concentrated solution of benzene at 277 K. IR : 3850 (vw), 3054 , 2955 , 1986 (s), 1821 , 1585 (vw), 1571 (vw), 1515 (vw), 1477 (w), 1434 (m), 1307 (vw), 1263 (vw), 1236 , 1182 (vw), 1159 (vw), 1119 (vw), 1092 (m), 1071 (vw), 1026 (vw), 997 (w), 907 (vw), 846 (vw), 741 (m), 690 (vs), 618 (vw), 565 (w), 541 (vw), 496 (m), 412 (vw).A few crystals of Pd2(CO)8(PPh3)2  were also isolated from this crystallization.Triphenylphosphine oxide  and Resp2), 0.98 Å (methyl) and 0.99 Å (methylene), with common isotropic temperature factors for all hydrogen atoms of the aromatic rings and methyl groups. SADI restraints on bond lengths and DELU restraints on anisotropic thermal parameters were used to model the disordered carbene ligand and diethyl ether molecule over two positions. For the refinement of 2, four reflections  global, 1, 2. DOI: 10.1107/S2056989021009270/tx20411sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989021009270/tx20412sup3.hklStructure factors: contains datablock(s) 2. DOI: 2108168, 2108167CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Intermolecular N—H⋯O and C—H⋯O hydrogen bonds connect these dimers to form a three-dimensional network. In addition, C—H⋯π interactions and π–π stacking interactions help to stabilize the packing.In the crystal, strong C—H⋯O and N—H⋯N hydrogen bonds form dimers with  19H17N3O2, adopts a screw-boat conformation. In the crystal, strong C—H⋯O and N—H⋯N hydrogen bonds form dimers with R22(14) and R22(12) ring motifs, respectively, between consecutive molecules along the c-axis direction. Intermolecular N—H⋯O and C—H⋯O hydrogen bonds connect these dimers, forming a three-dimensional network. C—H⋯π interactions and π–π stacking interactions contribute to the stabilization of the molecular packing. A Hirshfeld surface analysis indicates that the contributions from the most prevalent interactions are H⋯H (47.1%), C⋯H/H⋯C (20.9%), O⋯H/H⋯O (15.3%) and N⋯H/H⋯N (11.4%).The central tetrahydropyridine ring of the title compound, C The C7–C12 phenyl ring, which is attached to N1, is in an equatorial position and makes a dihedral angle of 54.43 (9)° with the mean plane of the tetrahydropyridine ring. The C13–C18 methoxyphenyl ring, which is attached to C4, is in an axial position. The dihedral angle between the C7–C12 phenyl and C13–C18 methoxyphenyl rings is 68.61 (10)°.The title compound Fig. 1 crystallC⋯N2 hydrogen bonds  −x + 1, −y + 1, −z + 1; (v) −x + y + z + Cg3 is the centroid of the C13–C18 methoxyphenyl ring; Table 1Cg2⋯Cg2iii = 3.8918 (15) Å and slippage = 1.551 Å; Cg2 is the centroid of the C7–C12 phenyl ring] contribute to the stabilization of the molecular packing  to 1.8469 (blue) a.u. The dnorm mapping indicates that strong hydrogen-bonding interactions, such as N—H⋯N, N—H⋯O and C—H⋯O hydrogen bonds , followed by C⋯H/H⋯C , O⋯H/H⋯O  and N⋯H/H⋯N . The percentage contributions of the C⋯C, C⋯N/N⋯C and N⋯N contacts are negligible, at 3.1, 1.4 and 0.8%, respectively. The predominance of H⋯H, C⋯H/H⋯C, O⋯H/H⋯O and N⋯H/H⋯N contacts indicate that van der Waals interactions and hydrogen bonding play the major roles in the crystal packing -6-oxo-1-phenyl-1,4,5,6-tetrahydropyridine-3-carbonitrile (I)  -2-(pyridin-2-yl)ethenyl]-1,4,5,6-tetrahydropyridine-3-carboxylate (III)   crystallizes in the monoclinic space group Pc with Z = 4, and with two molecules, A and B, in the asymmetric unit. These molecules are stereoisomers with an R,R absolute configuration at C3 and C4 in molecule A, whereas the corresponding atoms in B, C23 and C24, have an S configuration. In both molecules, the conformation of the central dihydropyridine ring is close to screw-boat. The molecular conformation is stabilized by N—H⋯O hydrogen bonds, forming a dimer with an c-axis direction. Furthermore C—Br⋯π and C=O⋯π stacking interactions between these ribbons contribute to the stabilization of the molecular packing.Compound (II) crystallizes in the monoclinic space group P21/c with Z = 4 and the asymmetric unit comprises one molecule. The central tetrahydropyridine ring is almost planar with a maximum deviation of 0.074 (3) Å for C4. The phenyl and dichlorophenyl rings are at an angle of 21.28 (15)°. They form dihedral angles of 86.10 (15) and 87.17 (14)°, respectively, with the central tetrahydropyridine ring. A strong intramolecular O2—H2⋯O1 hydrogen bond stabilizes the molecular conformation of the molecule, creating an S(6) ring motif. In the crystal, molecules are linked by intermolecular N—H⋯N and C—H⋯N hydrogen bonds, and N—H⋯π and C—H⋯π interactions, forming a three-dimensional network.Compound (III) , the cis configuration of the pyridinyl-vinyl fragment is stabilized by a strong intramolecular N—H⋯N hydrogen bond. The phenyl and pyridine rings are inclined to one another by 77.3 (1)°. In the crystal, inversion dimers are present via pairs of C—H⋯O hydrogen bonds and are further linked by C—H⋯O hydrogen bonds and C—H⋯π interactions.In molecule (IV) , the molecules form dimers by means of a pair of N—H⋯O hydrogen bonds. The 2(1H)-pyridone ring displays a screw-boat conformation.For compound (To a solution of 2-(4-methoxybenzylidene)malononitrile  and acetoacetanilide  in methanol (25 mL), 3-4 drops of piperidine were added and the mixture was stirred at 328–333 K for 10 min and was kept at room temperature for 48 h. Then 15 mL of methanol were removed from the reaction mixture, which was left overnight. The precipitated crystals were separated by filtration and recrystallized from ethanol/water (1:1) solution .1H NMR : 2.80 ; 3.19 ; 3.82 ; 3.93 ; 5.85 ; 7.15–7.58 . 13C NMR : 36.06 (CH—Ar), 40.42 (CH2), 53.78 (OCH3), 59.05 (Cquat.), 112.89 (2CHar), 121.21 (CN), 128.61 (CHar.), 128.88 (2CHar.), 130.44 (2CHar.), 130.51 (2CHar.), 136.06 (Car. quat.), 137.02 (Car. quat.), 154.59 (Car. quat.), 155.18 (Cquat.), 168.82 (N—C=O).C = 0.91 (2) and N3—H3D = 0.91 (2) Å with Uiso(H) = 1.2Ueq(N)]. C-bound H atoms were positioned geometrically, with C—H = 0.95–1.00 Å, and were refined with Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-methyl).Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S205698902200175X/vm2260sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902200175X/vm2260Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698902200175X/vm2260Isup3.cmlSupporting information file. DOI: 2152191CCDC reference:  crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:


This compound exists in the crystal phase as a methanol solvate.The molecular and crystal structures of the title compound, C 36H37ClN4O7·CH3OH, which crystallizes as a methanol solvate, may possess biological activity, which is inherent for a natural peptide or protein. In the crystal, molecules of the title compound form hydrogen-bonded tetramers with the solvate molecules acting as bridges as a result of the O—H⋯O and N—H⋯O intermolecular hydrogen bonds. Hirshfeld surface analysis was used to study the different types of intermolecular interactions whose contributions are: H⋯H = 53.8%, O⋯H/H⋯O = 19.0%, C⋯H/H⋯C = 14.8%, Cl⋯H/H⋯Cl = 5.3%, N⋯H/H⋯N = 3.2%.The title compound, C The target product contains a heterocyclic core bound to peptidomimetics, compounds that mimic a natural peptide or protein and which may have high biological activity. The pyrrolone fragment is also a privileged motif because of its biological activities, namely antibacterial  with the populations of A:B in a 0.303 (10):0.697 (10) ratio. All atoms of the partially saturated five-membered heterocycle are in the same plane with an accuracy of 0.008 Å. The N2—C4 bond length of 1.380 (3) Å and the C8—N2—C4—N1 torsion angle of 2.2 (4)° indicate conjugation between the π-systems of the partially saturated and oxazole cycles. The para-chlorophenyl substituent is located in the pseudo-equatorial position and is turned in relation to the C7—C8 endocyclic bond [C6—C7—C8—C9 = −120.9 (3)° and C7—C8—C9—C10 = 60.9 (3)°]. The C16(=O4)—N3 carbamide group is located in the -ac position in relation to the C6—C7 endocyclic bond [C6—C7—C15—C16 = −107.2 (3)°], and the C16=O4 carbonyl group is slightly non-coplanar to the C7—C15 bond [C7—C15—C16—O4 = 22.5 (4)°]. The para-methoxyphenyl substituent at the nitrogen atom is turned almost orthogonally to the plane of the carbamide group [C16—N3—C17—C22 = −99.5 (3)°]. The para-methoxyphenyl substituent at the carbon atom is located in a position intermediate between sp and −sc and is also rotated almost orthogonally to the plane of the carbamide group . In both para-methoxyphenyl substituents, the methoxy group is coplanar with the plane of the aromatic ring  despite the steric repulsion between the methyl group and the aromatic ring atoms . The substituent at the C24 atom is located in the −sc position relative to the N3—C16 bond [C32—C24—N3—C16 = −78.1 (3)°] and the C32—O6 carbonyl group is slightly non-coplanar to the N3—C24 bond [O6—C32—C24—N3 = −27.8 (3)°]. The tert-butyl substituent is located in antiperplanar position to the C32—C24 bond [C33—N4—C32—C24 = 172.9 (2)°].The title compound crystallizes as a methanol solvate Fig. 1. The metA⋯O8A, O3—H8B⋯O8B, O8A—H8A⋯O4 and O8B–H8B⋯O4 intermolecular hydrogen bonds . The highest contribution is from H⋯H contacts (53.8%), while these made by the O⋯H/H⋯O (19.0%) and C⋯H/H⋯C (14.8%) interactions are similar . The contributions of Cl⋯H/H⋯Cl (5.3%) and N⋯H/H⋯N (3.2%) interactions  are very small.In the two-dimensional fingerprint plots the pair of sharp spikes indicate strong hydrogen bonds and short contacts in the crystal structure Fig. 4a. The har Fig. 4c, 4d. Tns Fig. 4e, 4f aret al., 2016et al., 2019et al., 2018et al., 2003bet al., 2011et al., 2013et al., 2017et al., 2003aet al., 2011et al., 2009et al., 2015et al., 2005et al., 2019et al., 2019et al., 2008et al., 2016et al., 2018et al., 1991et al., 2006et al., 1990et al., 2016et al., 2001et al., 2019et al., 2004et al., 2014A search of the Cambridge Structural Database  (0.5 mmol) and 4-methoxybenzaldehyde (2) (0.5 mmol) were dissolved in methanol (4 mL) and stirred for 0.5 h. Then, 2-acetic acid (1) (0.5 mmol) and tert-butylisocyanide (4) (0.5 mmol) were added consistently and the reaction mixture was stirred for 24 h at 319 K. The mixture was allowed to stand overnight. The crystal precipitate was filtered off and dried. The reaction scheme is shown in Fig. 54-Methoxyaniline (Uiso(H) = 1.5Ueq for methyl and hydroxyl groups and with Car—H = 0.93 Å, Csp3—H = 0.97 Å, N—H = 0.89 Å Uiso(H) = 1.2Ueq for all other hydrogen atoms. The solvent molecule is disordered over two positions (A and B) in a 0.303 (10):0.697 (10) occupancy ratio. Csp3—O bonds were refined with fixed length of 1.413 Å.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989021011312/zq2268sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021011312/zq2268Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021011312/zq2268Isup3.cmlSupporting information file. DOI: 2118094CCDC reference:  crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:


The title compounds feature O—H⋯O carboxylic-acid inversion dimers and N—H⋯N chains in their extended structures. 11H11N3O3, (I), and C10H10N2O2, (II), are commercially available and were crystallized from ethyl acetate solution. The dihedral angle between the pyrazole and phenyl rings in (I) is 52.34 (7)° and the equivalent angle between the isoxazole and phenyl rings in (II) is 7.30 (13)°. In the crystal of (I), the molecules form carboxylic acid inversion dimers with an R(8) ring motif via pairwise O—H⋯O hydrogen bonds. In the crystal of (II), the molecules are linked via N—H⋯N hydrogen bonds forming chains propagating along [010] with a C(5) motif. A weak N—H⋯π interaction also features in the packing of (II). Hirshfeld surface analysis was used to explore the intermolecular contacts in the crystals of both title compounds: the most important contacts for (I) are H⋯H (41.5%) and O⋯H/H⋯O (22.4%). For (II), the most significant contact percentages are H⋯H (36.1%) followed by C⋯H/H⋯C (31.3%).The title compounds, C The C3—N3 distance of 1.353 (2) Å is typical for an amino group bound to an aromatic ring. The carboxylic carbon–oxygen distances are 1.255 (2) and 1.316 (2) for C4—O2 and C4—O1, respectively, indicating that the former bond may be affected by the intramolecular N—H⋯O hydrogen bond.The molecular structure of compound (I)d Table 1. This boif Fig. 1 with an Database survey section). The C3—N2 distance is 1.350 (5) Å and is typical of an amino group bound to an aromatic ring.The molecular structure of compound (II)via the O1—H1⋯O2i  link to generate an R(8) loop with O⋯O = 2.649 (2) Å, see Table 1via π–π interactions, notably weak stacking interactions between the 4-methoxyphenyl rings  along the a-axis direction.In the extended structure of (I)b-axis direction via the N2—H2A⋯N1i hydrogen bond , C6—H6⋯Cg1iii (2.86 Å) and C9—H9⋯Cg2ii (2.86 Å) . Analysis of the two-dimensional fingerprint plots indicates that the H⋯H (36.1%) interactions are the major factor in the crystal packing with C⋯H/H⋯C (31.3%) contacts making the next highest contribution. The percentage contributions of other weak interactions are: O⋯H/H⋯O (17.3%) and N⋯H/ H⋯N (12.1%). Figures showing the shape-index surface for each compound and the overall fingerprint plots are included in the supporting information.Fig. 6et al., 2016et al., 1974et al., 2007N-4-dimethyl-N-[3-{4-(trifluoromethyl)phenyl]-1,2-oxazol-5-yl}benzene-1-sulfonamide -1,2-oxazole -5-nitro-4,5-dihydro-1H-pyrazole-4-carboxylate Uiso(H) = 1.2Ueq(C) or 1.5Ueq(methyl C). In order to ensure a chemically meaningful O—H distance in (I)Uiso(H) = 1.5Ueq(O). In (I)Uiso(H) = 1.5Ueq(N). The absolute structure of (II)Crystal data, data collection, and structure refinement details are summarized in Table 310.1107/S2056989022001827/hb8013sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S2056989022001827/hb8013Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989022001827/hb8013IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989022001827/hb8013Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989022001827/hb8013IIsup5.cmlSupporting information file. DOI: 10.1107/S2056989022001827/hb8013sup6.pdfShape-index and fingerprint plots for compounds (I) and (II). DOI: 2152583, 2152582CCDC references:  crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:


Neighbouring (C 12H15N2)2[ZnCl4], is a salt with two symmetrically independent, essentially planar heterocyclic cations and a slightly distorted tetrahedral chlorozincate dianion. N—H⋯Cl hydrogen bonds link these ionic constituents into a discrete aggregate, which comprises one formula unit. The effect of hydrogen bonding is reflected in the minor distortions of the [ZnCl4]2− moiety: distances between the cation and chlorido ligands engaged in classical hydrogen bonds are significantly longer than the others. Secondary interactions comprise C—H⋯π hydrogen bonding and weak π–π stacking. A Hirshfeld surface analysis indicates that the most abundant contacts in packing stem from H⋯H (47.8%) and Cl⋯H/H⋯Cl (29.3%) interactions.The title compound, (C Mackinlaya sp.  tetrachloridozincate, an intermediate in the synthesis of mackinazolinone, using high-resolution diffraction data and Hirshfeld surface analysis.Tricyclic quinazolines are counted among the most exciting quinazoline alkaloids. Specifically, the alkaloid mackinazoline was isolated from P21/n space group, with two [C12H15N2]+ cations and a [ZnCl4]2− counter-anion in the asymmetric unit  are essentially coplanar, with r.m.s. deviations of 0.0437 and 0.0168 Å for molecules A and B, respectively. The remaining atoms of the third ring are significantly displaced above the opposite faces of these planes with deviations of 0.3877 (12) Å for C10A and 0.3831 (11) Å for C11A in residue A and 0.4705 (11) Å for C10B and 0.2495 (11) Å for C11B in residue B. Fig. 2The title compound crystallizes in the it Fig. 1. The benA and N1B, respectively, and the acquired positive charge is delocalized within the –N–C–N– moiety in the ring, where the N1A—C2A and N1B—C2B bonds are only slightly longer than C2A—N3A and C2B—N3B  fragment (see Table S1 in the supporting information).However, these C—N bond lengths are shorter than those in the related tricyclic protonated  and the very favourable ratio between observations and variables (100:1) in our diffraction data result in small standard uncertainties for atomic coordinates and derived geometric parameters and allow to discuss more subtle details. The most acute angle of 103.33 (11)° within the tetrachloridozincate dianion  graph-set motif  Å] and involves an interaction between the electron-rich equatorial region of the halogen atom and the ring atom attached to two N-atom neighbours, most probably the most electron-deficient atom in the heterocycle. These contacts link anions and cations into a three-dimensional network. Weak π–π stacking interactions occur between pyrimidine  and benzene  rings of antiparallel pairs of cations and involve contact distances of Cg1⋯Cg3  = 3.6225 (5) Å (slippage 0.857 Å) and of Cg7⋯Cg9  = 3.6246 (7) Å (slippage 0.994 Å).The crystal packing is further stabilized by intermolecular C—H⋯π interactions Table 2 and addiCrystalExplorer17.5  analysis  in Fig. 6ly Fig. 6. Minor cet al., 20164]2− anions. They are associated with refcodes PODLUP  of 2,3-tetramethylenquinazoline-4-one Fig. 7 were plaUiso(H) = 1.2Ueq(C). H atoms bonded to nitrogen were located in a difference-Fourier map, and their positional and isotropic displacement parameters were freely refined.Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989021004989/jq2006sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021004989/jq2006Isup3.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021004989/jq2006sup4.docxSupporting information file. DOI: 2082994CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Both polymorphs are monoclinic and the [Rh(μ-I)(COD)]2 molecules in each case possess C2v symmetry. However, the core geometry of the butterfly-shaped Rh2I2 core differs substantially. In the C2/c polymorph, the core geometry of [Rh(μ-I)(COD)]2B is bent, with a hinge angle of 96.13 (8)° and a Rh⋯Rh distance of 2.9612 (11) Å. The P21/c polymorph features a more planar [Rh(μ-I)(COD)]2P core geometry, with a hinge angle of 145.69 (9)° and a Rh⋯Rh distance of 3.7646 (5) Å.The solid-state structure of di-μ-iodido-bis{rhodium(I)}, [Rh M(μ-X)(COD)]2,  are ubiquitous synthons for Rh and Ir catalysts. As a representative example, [Rh(COD)(DPEphos)]BF4 catalyzes the hydroamination of vinylarenes with anti-Markovnikov selectivity, and is prepared via the reaction of [Rh(μ-Cl)(COD)]2 with two equivalents of AgBF4 and two equivalents of DPEphos (COD)]2 , all compounds have been structurally characterized with the notable exception of [Rh(μ-I)(COD)]2 (COD)]2 containing dimers with significantly different structural features. X-ray diffraction of the dark-orange crystals revealed that [Rh(μ-I)(COD)]2 crystallized in the monoclinic C2/c space group in this polymorph. The molecular structure of [Rh(μ-I)(COD)]2B exhibited a C2v-symmetric geometry featuring a bent Rh2I2 diamond core (COD)]2 and [Rh(μ-Br)(COD)]2, which both exhibit more planar Rh2X2 cores with hinge angles of 169.08 (6) and 148.74 (7)° and Rh⋯Rh distances of 3.5169 (6) and 3.5648 (14) Å, respectively (COD)]2 re Fig. 1. The hin2, this time crystallizing in the monoclinic P21/c space group. The geometry of the dirhodium dimer in the P21/c polymorph [Rh(μ-I)(COD)]2P differs significantly from the C2/c polymorph [Rh(μ-I)(COD)]2B. The Rh2I2 core geometry of [Rh(μ-I)(COD)]2P is more planar, with a hinge angle of 145.69 (9)° and a Rh⋯Rh distance of 3.7646 (5) Å (COD)]2 and [Rh(μ-Br)(COD)]2 (COD)] Å Fig. 2. The mol−1) between planar and bent [Rh(μ-X)(L)2]2 geometries (COD)]2B but not [Rh(μ-Cl)(COD)]2 and [Rh(μ-Br)(COD)]2 (COD)]2.A previous theoretical study found relatively small energetic differences  and 2.6975 (7) Å]. The four Rh—I distances in [Rh(μ-I)(COD)]2P are slightly less symmetric: the bonds between I2 and the two Rh centers [2.6833 (4) and 2.6738 (4) Å] are slightly shorter than those associated with I1 [2.6998 (4) and 2.7061 (4) Å]. Similarly, the Rh—C distances in [Rh(μ-I)(COD)]2B are more symmetric, ranging from 2.115 (6) to 2.122 (6) Å, while the Rh—C distances in [Rh(μ-I)(COD)]2P range from 2.117 (4) to 2.131 (4) Å. The average Rh—C distance in the bent and planar structures are similar to the average Rh—C distances reported for the [Rh(μ-Cl)(COD)]2 and [Rh(μ-Br)(COD)]2 analogues (COD)]2P  are considerably longer than the average Rh—Br and Rh—Cl distances in [Rh(μ-Br)(COD)]2 and [Rh(μ-Cl)(COD)]2 .There is no meaningful difference between the two independent Rh—I distances in [Rh(μ-I)(COD)]2 are attributed to differences in crystal packing and weak interatomic forces. The bent and planar geometries are likely similar in energy. Stabilization of the bent geometry in [Rh(μ-I)(COD)]2B arises from intramolecular dispersion forces between the C—H bonds of the cyclooctadiene ligands on the two Rh centers within each molecule. Indeed there are four close C—H⋯H—C contacts  between the alkene and methylene hydrogen atoms made possible by the bent geometry (COD)]2P are two Rh⋯H—C contacts in the apical positions of Rh2  = 2.67 Å; Rh2⋯H3A = 2.93 Å) that could, at best, be labeled as weak intermolecular agostic interactions  than the bent C2/c polymorph (2.597 g cm−3), indicative of tighter crystal packing.The structural differences between the dimers in the two polymorphs of [Rh(μ-I)(COD)]ry Fig. 3. In the ns Fig. 4. Consist2 was prepared according to the procedure described by J. A. Hlina et al.  was added to toluene (5 mL) and trimethylsilyl iodide . The reaction mixture turned dark red and rust-colored crystals precipitated from the solution. The solid was isolated, washed with hexanes and dried in vacuo to yield the final product [Rh(μ-I)(COD)]2 as a red–brown crystalline solid . X-ray quality crystals were grown from a concentrated solution of toluene at 236 K, resulting in crystals with two different morphologies. 1H NMR (C6D6): 1.15–1.28 , 1.90–2.05 , 4.62–4.70 . A single species was observed by 1H NMR spectroscopy and the 1H NMR spectrum did not contain any broad features indicative of dynamic behavior or interconversion between the two isomers in solution.[Rh(μ-I)(COD)] al. 2017. Under aSHELXT) produced a complete phasing model for refinement. All non-hydrogen atoms were refined anisotropically by full-matrix least-squares (SHELXL2018). All carbon-bonded methylene hydrogen atoms were placed using a riding model. Their positions were constrained relative to their parent atom using the appropriate HFIX command in SHELXL2018. All carbon-bonded methine hydrogen atoms were located in the difference map. Their C—H distances were restrained to a target value of 1.00 (2) Å. For all H atoms, displacement parameter Uiso(H) values were set to 1.2 times Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table 110.1107/S205698902100743X/zl5015sup1.cifCrystal structure: contains datablock(s) global, B, P. DOI: 10.1107/S205698902100743X/zl5015Bsup2.hklStructure factors: contains datablock(s) B. DOI: 10.1107/S205698902100743X/zl5015Psup3.hklStructure factors: contains datablock(s) P. DOI: 2097568, 2097567CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Saccharomyces cerevisiae is its ability to perform simultaneous respiration and fermentation at high growth rate even under fully aerobic conditions. In the present work, this Crabtree effect called phenomenon was investigated in detail by comparative 13C metabolic flux analysis of S. cerevisiae growing under purely oxidative, respiro-fermentative and predominantly fermentative conditions.One of the most fascinating properties of the biotechnologically important organism S. cerevisiae exhibited a by-pass of pyruvate dehydrogenase in all physiological regimes. During oxidative growth this by-pass was mainly provided via pyruvate decarboxylase, acetaldehyde dehydrogenase, acetyl-CoA synthase and transport of acetyl-CoA into the mitochondrion. During fermentative growth this route, however, was saturated due to limited enzyme capacity. Under these conditions the cells exhibited high carbon flux through a chain of reactions involving pyruvate carboxylase, the oxaloacetate transporter and malic enzyme. During purely oxidative growth the PPP alone was sufficient to completely supply NADPH for anabolism. During fermentation, it provided only 60 % of the required NADPH.The metabolic shift from oxidative to fermentative growth was accompanied by complex changes of carbon flux throughout the whole central metabolism. This involved a flux redirection from the pentose phosphate pathway (PPP) towards glycolysis, an increased flux through pyruvate carboxylase, the fermentative pathways and malic enzyme, a flux decrease through the TCA cycle, and a partial relocation of alanine biosynthesis from the mitochondrion to the cytosol. S. cerevisiae possesses different metabolic by-passes to channel carbon into the mitochondrion. This involves the conversion of cytosolic pyruvate either into acetyl CoA or oxaloacetate followed by intercompartmental transport of these metabolites. During oxidative growth mainly the NAD specific isoforms of acetaldehyde dehydrogenase and isocitrate dehydrogenase catalyze the corresponding reactions in S. cerevisiae, whereas NADPH supply under fermentative conditions involves significant contribution of sources other than the PPP such as e. g. NADPH specific acetaldehyde dehydrogenase or isocitrate dehydrogenase.We conclude that, in order to overcome the limited capacity of pyruvate dehydrogenase,  S. cerevisiae is an important biotechnological organism e.g. for production of ethanol [S. cerevisiae is able to perform simultaneous respiration and fermentation at high growth rates even under fully aerobic conditions [S. cerevisiae by a shift from biomass formation towards fermentative products at increased dilution rates [13C metabolic flux analysis which allows the quantification of in vivo activity of pathways and reactions [S. cerevisiae and related yeast strains [S. cerevisiae by quantifying metabolic fluxes through the underlying reactions in the central metabolism at these different physiological states. For this purpose chemostat cultures with [1-13C] glucose as substrate were grown at different growth rates under aerobic glucose-limited conditions to metabolic and isotopic steady-state. The use of a continuous culture hereby allowed the precise adjustment of the physiological state of the cells, i.e. the relative activity of the fermentative pathways.The budding yeast  ethanol , recombi ethanol , antibio ethanol  or bioma ethanol . Additio ethanol ,6. This nditions ,8. This on rates . Despiteon rates -12, a clon rates ,14. The on rates ,16. A poeactions -19. In r strains -27. In t-1 and 0.45 h-1 , mixed respiro-fermentative (0.30 h-1), and mainly fermentative metabolism (0.40 h-1) were selected for the flux studies. The resulting ethanol yield under these conditions was 0.0, 0.5, and 1.1 mol ethanol (mol glucose)-1, respectively. The stoichiometric data obtained from the 13C cultures agreed well with the corresponding values from the initially performed cultivations  The metabolic shift had a strong effect on the flux partitioning between glycolysis and PPP. Whereas, at purely oxidative metabolism, the major carbon flux was channeled into the PPP, the opposite was observed for fermentative metabolism. The data of the present work together with previous flux studies of t Figure . A simil 0.4 h-1 . The red 0.4 h-1 . Due to S. cerevisiae  glucose . Cultivations were performed in a continuously operated bioreactor  with a culture volume of 100 ml at 30°C and 500 rpm. The aeration rate was kept at 100 mL/min by a mass flow controller . Composition of inlet and exhaust gas was determined on-line by mass spectrometry as described previously  was obtained. Enzymatic test-kits were used for quantification of glucose, glycerol, acetate  and ethanol  in the supernatant. Concentrations of 2-oxoglutarate, pyruvate, succinate, and fumarate in the supernatant were analyzed by HPLC [S. cerevisiae was quantified by HPLC [i.e. the quantification of the yield coefficients, was 3.7 % (biomass yield), 4.0 % (ethanol yield), 9.3 % (acetate yield) and 11.3 % (glycerol yield).Optical density (OD by HPLC  after hy0) to the fully 13C labeled (xn) variant could be measured with good signal intensity and accuracy  is shown in Figure S. cerevisiae [The metabolic network of revisiae . For glyrevisiae ,46 was t1 - ν2 - ν3 + ν4 - ν5 = 0     (1)Glucose 6-phosphate: ν3 - ν4 - ν6 + ν8 - ν9 + ν10 - ν11 = 0     (2)Fructose 6-phosphate: ν2 - ν7 - ν8 + ν9 - 2ν12 + 2ν13 = 0     (3)Pentose phosphate: ν8 + ν9 + ν10 - ν11 - ν14 = 0     (4)Erythrose 4-phosphate: - ν12 - ν13 - ν10 + ν11 - ν5 = 0     (5)Sedoheptulose 7-phosphate: ν6 - ν10 + ν11 + ν8 - ν9 + ν12 - ν13 + ν16 - ν17 - ν18 = 0     (6)Glyceraldehyde 3-phosphate: ν6 - ν15 - ν16 = 0     (7)Dihydroxyacetone-phosphate: ν17 - ν19 - ν20 - ν24 = 0     (8)3-Phosphoglycerate: ν20 - ν21 - ν22 = 0     (9)Serine: ν22 - ν23 + ν29 = 0     (10)Glycine: ν28 - ν29 - ν30 = 0     (11)Threonine: ν24 - ν25 - ν26 = 0     (12)Phosphoenolpyruvate: νcyt: ν26 - ν27 - ν32 - ν33 - ν34 = 0     (13)Pyruvate32 - ν37 - ν38 = 0     (14)Acetaldehyde: ν38 - ν39 - ν40 = 0     (15)Acetate: νcyt: ν40 - ν41 - ν47 + ν48 = 0     (16)Acetyl CoAcyt: ν33 - ν42 = 0     (17)Alaninecyt: ν27 - ν28 - ν31 - ν35 + ν36 = 0     (18)Oxaloacetatemit: ν34 - ν43 - ν44 + ν50 - ν51 = 0     (19)Pyruvatemit: ν43 - ν45 = 0     (20)Alaninemit: ν44 - ν46 + ν47 - ν48 - ν49 = 0     (21)Acetyl CoA49 - ν52 - ν53 = 0     (22)2-Oxoglutarate: ν53 - ν54 + ν55 = 0     (23)Succinate: νmit: ν35 - ν36 - ν49 - ν50 + ν54 - ν55 = 0     (24)Oxaloacetate/MalateS. cerevisiae was considered  combining metabolite balancing and isotopomer modeling. The performed approach utilized metabolite balancing  during each optimization step considering stoichiometric data on anabolic demand for biomass precursors Table  and prod13C tracer studies and all analytics involved.OF carried out the experimental work including the chemostat cultivations, the S. cerevisiae, the software implementation, the calculation of carbon fluxes, and the statistical analysis. He drafted and composed the manuscript.CW designed the study and carried out all computational work involving the construction of the metabolic network model of Both authors read and approved the final manuscript.


CENP‐A nucleosomes. Centromere maintenance during the cell cycle requires HJURP‐mediated CENP‐A deposition, a process regulated by the Mis18 complex (Mis18α/Mis18β/Mis18BP1). Spatial and temporal regulation of Mis18 complex assembly is crucial for its centromere association and function. Here, we provide the molecular basis for the assembly and regulation of the Mis18 complex. We show that the N‐terminal region of Mis18BP1 spanning amino acid residues 20–130 directly interacts with Mis18α/β to form the Mis18 complex. Within Mis18α/β, the Mis18α MeDiY domain can directly interact with Mis18BP1. Mis18α/β forms a hetero‐hexamer with 4 Mis18α and 2 Mis18β. However, only two copies of Mis18BP1 interact with Mis18α/β to form a hetero‐octameric assembly, highlighting the role of Mis18 oligomerization in limiting the number of Mis18BP1 within the Mis18 complex. Furthermore, we demonstrate the involvement of consensus Cdk1 phosphorylation sites on Mis18 complex assembly and thus provide a rationale for cell cycle‐regulated timing of Mis18 assembly and CENP‐A deposition.The centromere, a chromosomal locus that acts as a microtubule attachment site, is epigenetically specified by the enrichment of  Equal and identical distribution of chromosomes to each daughter cell during cell division is essential for maintaining genome integrity. A central player regulating this process is the kinetochore, a large proteinaceous structure assembled at a specialized region of the chromosome called the centromere Schizosaccharomyces pombe); however, the molecular mechanisms by which it is achieved share considerable similarity In most eukaryotes, the centromeric chromatin is epigenetically defined by the enrichment of nucleosomes containing the histone H3 variant CENP‐A S. pombe and humans The Mis18 proteins contain two structurally distinct domains , both of which can self‐oligomerize. Previously, we and others have shown that oligomerization of Mis18 proteins is required for their centromere association and function both in 20–130), binds Mis18α/β hetero‐oligomer via the Mis18α/β MeDiY hetero‐dimer to form the Mis18 complex. Characterization of the oligomeric structures of the Mis18 subcomplexes revealed that the Mis18α/β complex is a hetero‐hexamer with four copies of Mis18α and two copies of Mis18β, while the Mis18 holo‐complex is a hetero‐octamer with two additional copies of Mis18BP1. We identify two conserved consensus Cdk1 phosphorylation sites (T40 and S110) within Mis18BP120–130 and show that phospho‐mimicking mutations of these residues disrupt Mis18 complex formation. This explains the molecular basis for Cdk1‐mediated timing of Mis18 assembly and CENP‐A deposition.Here, we show that Mis18BP1, through its highly conserved N‐terminal region comprising amino acids 20–130  integrated in a chromosome arm fl) as well as two different N‐terminal fragments: mCherry‐Mis18BP120–130 and mCherry‐Mis18BP1336–483 (covering the SANTA domain) to the tethering site. Mis18α tethered to the ectopic alphoidtetO array recruited Mis18BP1fl and Mis18BP120–130 more robustly compared to the Mis18BP1336–483  and SANT (residues 878–927) domains Figs A. It has–130 Fig B. These 20–130 can directly interact with Mis18α/β in vitro, individually purified recombinant proteins (Mis18α/β and Mis18BP120–130) were analyzed using size‐exclusion chromatography (SEC) before and after complex formation. While Mis18α/β and Mis18BP120–130 eluted at 11.0 and 15.7 ml, respectively, the complex containing Mis18α/β and Mis18BP120–130 eluted at 10.9 ml  and a C‐terminal α‐helical domain  and Mis18BP120–130 (12.7 kDa) migrate similarly in the SDS–PAGE . While His‐GFP‐Mis18αMeDiY on its own eluted at 13.8 ml, the His‐GFP‐Mis18αMeDiY/Mis18BP120–130 complex eluted at 13.3 ml  showed that they elute at distinct elution volumes, 13.9 and 12.2 ml, respectively, demonstrating a lack of interaction . Consistent with the SEC analysis shown in Figs MeDiY could bind Mis18BP120–130. His‐GFP‐Mis18βMeDiY failed to interact with Mis18BP120–130 even under a low ionic strength condition . The measured MW of the untagged Mis18α/β complex was 151.2 ± 2.9 kDa  and correlated well with the calculated MW of a hetero‐hexamer , it is almost impossible to accurately determine the subunit stoichiometry within the Mis18 hetero‐hexamer. Hence, we introduced a noticeable size variation by purifying His‐GFP‐Mis18α in a complex with His‐Mis18β. Interestingly, the measured MW of His‐GFP‐Mis18α/His‐Mis18β complex was 287.2 ± 5.5 kDa, matching the calculated MW of a hetero‐hexamer containing four copies of His‐GFP‐Mis18α and two copies of His‐Mis18β  and His‐MBP‐Mis18βC‐term (Mis18β 184‐end), we reconstituted the C‐terminal helical assembly and analyzed their composition using SEC‐MALS  by SEC‐MALS (175.4 ± 3.3 kDa) is in agreement with the calculated MW of a hetero‐octamer containing a Mis18α/β hetero‐hexamer plus two copies of Mis18BP120–130  matches the calculated MW  at the MeDiY dimeric interface of Mis18α/β and L136A/Y152A/C167A/S169K (Mis18αPocketM) in the putative substrate‐binding pocket using a S. pombe‐based homology‐modeled structure , His‐GFP‐Mis18αDimerM/His‐Mis18β (Mis18αDimerM/β), and His‐GFP‐Mis18αPocketM/His‐Mis18β (Mis18αPocketM/β) complexes using a Ni‐NTA pull‐down assay. While Mis18αwt/β and Mis18αPocketM/β interacted with Mis18BP120–130, Mis18αDimerM/β failed to do so efficiently . This strengthens the notion that the Mis18α/β hetero‐trimer is the core oligomeric unit formed through the interactions of the C‐terminal helices which then assemble into a hetero‐hexamer via the MeDiY‐dimerization interface  or in combination (Mis18BP120–130 T40E/S110D), on Mis18 complex formation using SEC. Both Mis18BP120–130 T40E and Mis18BP120–130 S110D interacted with Mis18α/β. However, Mis18BP120–130 T40E/S110D failed to form a complex with Mis18α/β  was recruited by TetR‐eYFP‐Mis18α to the array throughout the cell cycle. Consistent with this, recombinantly purified Mis18BP120–130 T40A/S110A interacted with Mis18α/β as analyzed by SEC . While mutating both these amino acid residues to phospho‐mimicking residues completely abolished the ability of Mis18BP1 to bind Mis18α/β in vitro and in vivo, individual phospho‐mimic mutants (T40E or S110D) failed do so efficiently. This suggests that Mis18BP1 binds Mis18α/β possibly via a bipartite binding mode. Overall, our findings together with the recent independent study by Pan et alThe temporal regulation of Mis18 complex formation defines the timing of HJURP‐mediated CENP‐A deposition essential for centromere inheritance and function imer Fig A. HoweveMeDiY, Mis18αC‐term, Mis18β, Mis18βMeDiY, and Mis18βC‐term were amplified from codon‐optimized sequences (GeneArt) while Mis18BP120–130 was amplified from a human cDNA library . Amplifications were then cloned into pET His6 msfGFP TEV, pET His6 TEV, pET His6 Sumo TEV, pET His6 MBP TEV, pGEX‐6P‐1 , pEC‐K‐3C‐His‐GST, and pEC‐K‐3C‐His LIC vectors. pET His6 msfGFP TEV (9GFP Addgene plasmid # 48287), pET His6 TEV (9B Addgene plasmid # 48284), pET His6 Sumo TEV (14S Addgene plasmid # 48291), and pET His6 MBP TEV (9C Addgene Plasmid #48286) were a gift from Scott Gradia.Human Mis18α, Mis18αDimerM; V82E/Y176D) and putative substrate‐binding pocket mutant  and Mis18BP1 mutants, Mis18BP120–130 T40E, Mis18BP120–130 S110D, Mis18BP120–130 T40E/S110D, and Mis18BP120–130 T40A/S110A were generated following the Quikchange site‐directed mutagenesis protocol (Stratagene).Mis18α MeDiY dimer‐disrupting mutant (Mis18αE. coli BL21 gold grown in LB media. Cultures were induced with 0.35 mM IPTG at 18°C overnight. Cell lysis was carried out by sonicating cells re‐suspended in a lysis buffer containing 20 mM Tris (pH 8.0), 250 mM NaCl (or 500 mM for Mis18BP120–130), 35 mM imidazole, and 2 mM βME. Lysis buffer was supplemented with 10 μg/ml DNase, 1 mM PMSF, and cOmplete . Proteins were purified from the clarified lysates by affinity chromatography using a 5 ml HisTrap HP column . The protein‐bound resin was washed with lysis buffer, followed by a buffer containing 20 mM Tris (pH 8.0), 1 M NaCl, 50 mM KCl, 10 mM MgCl2, 2 mM ATP, 35 mM imidazole, 2 mM βME, and with a final lysis buffer wash. Proteins were eluted using a lysis buffer supplemented with 500 mM imidazole and dialyzed overnight into 20 mM Tris (pH 8.0), 75–100 mM NaCl, and 2 mM DTT. All proteins were subjected to anion exchange chromatography using the HiTrap Q column . Appropriate fractions were pooled, concentrated, and injected into a Superdex 200 increase 10/300 or Superdex 75 10/300 column  equilibrated with 20 mM Tris (pH 8.0), 100–250 mM NaCl, and 2 mM DTT. Fractions were analyzed on SDS–PAGE and stained with Coomassie blue.Proteins were expressed, either individually or together with their binding partners, using 20–130 and made up to 200 μl with buffer before incubated for 30 min at 4°C with 60–120 μl of slurry that had been equilibrated in buffer. Beads were then washed four times with 1 ml of buffer, and bound protein was eluted by boiling in SDS–PAGE loading dye before being analyzed by SDS–PAGE.Ni‐NTA pull‐down assay was performed in 20 mM Tris (pH 8.0), 75–500 mM NaCl, 10% glycerol, 0.5% NP‐40, 35 mM imidazole, and 2 mM βME. Proteins were mixed with two times molar excess of Mis18BP1280 nm were analyzed by a homo‐polymer model  using the following parameters for: ∂A280 nm/∂c = 0.97 AU ml mg−1 (Mis18α/β complex), 0.86 AU ml mg−1 (His‐GFP‐Mis18α/His‐Mis18β and His‐Mis18α/His‐GFP‐Mis18β), 0.82 AU ml mg−1 (Mis18α/Mis18β/Mis18BP120–130), 0.71 AU ml mg−1 (Mis18α/Mis18β/His‐Sumo‐Mis18BP120–130), 1.10 AU ml mg−1 (His‐GFP‐Mis18αC‐term/His‐MBPMis18βC‐term), 0.86 AU ml mg−1 (His‐GFP‐Mis18α2M/His‐Mis18β), ∂n/∂c = 0.185 ml g−1, and buffer RI value of 1.335. The mean standard error in the mass accuracy determined for a range of protein–protein complexes spanning the mass range of 6–600 kDa is ±1.9%.Size‐exclusion chromatography  coupled to UV, static light scattering, and refractive index detection  was used to determine the absolute molecular mass of proteins and protein complexes in solution. Injections of 100 μl of about 1 mg/ml material were run on a Superdex 200 10/300 GL  size‐exclusion column pre‐equilibrated in 50 mM HEPES (pH 8.0), 150 mM NaCl, and 1 mM TCEP at 22°C with a flow rate of 0.5 ml/min. Light scattering, refractive index (RI), and Afl, Mis18BP120–130, and Mis18BP1336–483 were amplified from a human cDNA library  and cloned into the pCDNA3‐mCherry or ‐mCerulean LIC cloning vectors . Mis18α was cloned into the TetR‐eYFP‐IRES‐Puro vector. Mutant constructs, TetR‐eYFP‐Mis18αDimerM, mCherry‐Mis18BP120–130 T40E, mCherry‐Mis18BP120–130 S110D, mCherry‐Mis18BP120–130 T40E/S110D, and mCherry‐Mis18BP120–130 T40A/S110A, were generated using the Quikchange mutagenesis protocol (Stratagene).Human Mis18β, Mis18BP1tetO array) integrated in a chromosome arm, was maintained in DMEM (Gibco) supplemented with 5% fetal bovine serum (Biowest) and penicillin/streptomycin (Gibco). Cells were grown at 37°C and 5% CO2. Transfections were performed in parallel with XtremeGene‐9 (Roche) following manufacturer's instructions. Briefly, 24 h after plating, cells grown in 12‐well plates were incubated with transfection complexes containing: 0.25 μg of each vector, 100 μl of Opti‐MEM (Invitrogen), and 3 μl of XtremeGene‐9 reagent for 36 h.The HeLa 3‐8 overexpressing CENP‐A‐SNAP integration cell line, containing a synthetic α‐satellite  DNA array integration with tetO sites  for 5 min at room temperature and blocked with permeabilization buffer containing 3% BSA for 1 h at 37°C. Mouse anti‐CENP‐A  and Cy‐5‐conjugated donkey anti‐mouse secondary antibody (Jackson Immunoresearch) were used for centromere staining.z‐stacks were deconvolved using SoftWorx and analyzed using ImageJ software . For CENP‐A signal quantification, a custom‐made macro in ImageJ modified from Bodor et alTetO array  was determined for every z‐section within a 7‐square pixel box. The mean signal intensity in the array was obtained and the background subtracted using the minimum intensity values within the section. The average intensity of the CENP‐A signal from endogenous centromeres was used to normalize. At least three biological independent experiments were performed for each assay.Micrographs were acquired in the Centre Optical Instrumentation Laboratory on a DeltaVision Elite system (Applied Precision) using an inverted Olympus IX‐71 stand, with an Olympus UPlanSApo 100× oil immersion objective  1.4) and a Lumencor light source. Camera (Photometrics Cool Snap HQ), shutter and stage were controlled through SoftWorx (Applied Precision). The 0.2‐μm‐spaced AAJ conceived the project. FS, BM‐P, MAA, OM, AAJ, and WCE designed experiments. FS, BM‐P, MAA, MAW, and OM performed experiments and analyzed data. FS, BM‐P, MAA, OM, WCE, and AAJ wrote the manuscript.The authors declare that they have no conflict of interest.Expanded View Figures PDFClick here for additional data file.Review Process FileClick here for additional data file.


The crystal structure of 2-(5-bromothiophen-2-yl)acetonitrile, previously reported as a liquid, has short centrosymmetric Type I Br⋯Br halogen interactions. 6H4BrNS, crystallizes in the space group P21/n with one complete molecule in the asymmetric unit. The non-H atoms are nearly planar (r.m.s for non-H atoms = 0.071 Å), with the nitrile group oriented antiperiplanar with respect to the thiophene S atom. Intermolecular Type I centrosymmetric Br⋯Br halogen interactions are present at a distance of 3.582 (1) Å and with a C—Br⋯Br angle of 140.7 (1)°. Additional weaker C—H⋯N, C—H⋯S, and S⋯π interactions are also present. A Hirshfeld analysis indicates Br⋯Br interactions comprise only 1.9% of all the interatomic contacts.The title compound, C The large number of reflections in the data sets (and the Fourier-transform relationship of intensities to atoms) ensures that no particular bias was thereby introduced.10.1107/S2056989018000968/dx2004sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989018000968/dx2004Isup2.hklStructure factors: contains datablock(s) I. DOI: 1817195CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


In the two structures, short cationic tristannatoxanes (C 3(CH3)9(OH)2(H2O)2][Sn(CH3)3(CHO2)2] (1) and [Sn3(CH3)9(OH)2]Cl·H2O (2), are partially condensed products of hydrolysed trimethyltin chloride. In the structures of 1 and 2, short cationic tristannatoxanes (C9H29O2Sn3) are bridged by a diformatotrimethyltin anion or a chloride anion, respectively. Hydrogen bridges are present and supposedly stabilize these structures against further polymerization to the known polymeric trimethyltin hydroxide. Especially noteworthy is that the formate present in this structure was formed from atmospheric CO2.The title compounds, [Sn Therefore, the activation of CO2 plays an important role in today’s research. It is already known that CO2 is activated by electroreduction of different metals ° while in 2 it is 135.30 (17)°. In the chloride structure 2, a change in two further angles is noticed. The O1—Sn1—Cl1 angle [177.58 (10)°] and the O2—Sn3—Cl1′ angle [175.5 (12)°] decreases  and the chloride anion.In the crystal structures, no polymeric Sn–O structures were formed, as found in the trimethyltin hydroxide. The short trimethyltin hydroxide chain has a positive and the chloride or bisformatostannate a negative charge. In the structure of via hydrogen bonds. In structure 1 , thus forming a two-dimensional network. Additionally, hydrogen bonds between these sheets form a two-dimensional network along the bc plane (O4⋯H1—O1).As described, both structures are intermolecularly linked  1 Fig. 3, the for2  Å], one by a Cl1⋯H1i—O1i hydrogen bond [3.251 (4) Å]. A fourth hydrogen bond, Cl1⋯H3ii—O3ii [3.068 (5) Å], results in a distorted tetrahedral environment. Thus, a three-dimensional network of hydrogen bridges is formed. The interactions between Sn–Cl differ due to steric repulsion of the C2 and C7iii methyl groups. The van der Waals radius of a methyl group is 2 Å ° for Sn1—Cl1—Sn2 and 179.2 (2)° for O1—Sn1—Cl1 were observed  = 1.5Ueq(Cmethyl). The CH3 hydrogen atoms were allowed to rotate but not to tip. Due to point group symmetry 2 of both the cation and anion in 1, with the twofold rotation axis running through the respective central Sn atom and one of the methyl groups, the latter is equally disordered over two positions.Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989016014912/su5326sup1.cifCrystal structure: contains datablock(s) Global, 1, 2. DOI: 10.1107/S2056989016014912/su53261sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989016014912/su53262sup3.hklStructure factors: contains datablock(s) 2. DOI: 1505529, 1505528CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Two new crystal structures of eight- and ten-membered cyclic bisanisylphosphonothioyl disulfanes have been determined and these are compared to the structures of their ferrocenyl analogues. 5,5λ5-dioxadithiadiphosphocane-2,5-dithione, C16H18O4P2S4, and 2,5-bis(4-methoxyphenyl)-1,6,3,4,2λ5,5λ5-dioxadithiadiphosphecane-2,5-dithione, C18H22O4P2S4, have been determined and compared to structures of the ferrocenyl analogues. The eight-membered rings have similar conformations (TBC) but the ten-membered macrocycles are differently puckered. Structural parameters of the relevant SPSSPS motif have been analysed and are discussed in detail. Compound 1 was refined as an inversion twin and 2 was refined as a two-component rotational twin.Two new crystal structures of eight- and ten-membered cyclic bisanisylphosphonothioyl disulfanes, namely 2,5-bis(4-methoxyphenyl)-1,6,3,4,2λ Lawesson reagent LR P(S)SH and Fc(RO)P(S)SH, obtained in a simple reaction between LR or fLR and alcohols, are of considerable interest because they form a plethora of structurally interesting chelate complexes with metal ions S]e.g. eight-membered 1a  acid derivatives and among them the unique eight-, nine- and ten-membered cyclic bisanisylphosphonothioyl disulfanes . The related ferrocenyl compound 1a crystallizes in space group C2/c with non-typical three and half independent molecules in the asymmetric unit (Z = 28), which complicates comparisons.Views of molecular structures and atom-labeling scheme for 2 forms a monoclinic crystalline phase obeying P21/c space-group symmetry with one molecule in the asymmetric unit and Z = 4. The related ferrocenyl structure 2a crystallizes in space group PCompound trans arrangement, i.e. above and below the macrocycle ring plane for all compounds 1–2a, which is also typical for all open-chain bisphosphorothioyl disulfanes studied previously , 2.0704 (10), 2.0685 (10), 2.0711 (15) Å for 1a; 2.074 (3) Å for 2 and 2.0788 (9) Å for 2a. They are longer than the typical S—S bond lengths for known diorganyl disulfanes RSSR [2.05 (3) Å]. The observed S—S bond elongation in 1–2a may be correlated with the PSSP torsion angles  are shorter than the endocyclic P—S bonds (ca 2.10 Å).The S—S bond lengths have values of 2.068 (2) Å for 1–2a adopt a distorted tetrahedral geometry, where the C—P=S angles deviated the most (116.1–118.5°) from the ideal tetrahedral angle. This is obviously due to the steric effects of the anisyl and ferrocenyl substituents. On the other hand, it is worthy to note that the O—P–S bond angles in 1–2a (107–108°) are not distorted, probably due to minimal conformational strain present in those medium-sized heterocycles. Moreover, both the P=S and aromatic anisyl groups in 1 are almost perfectly coplanar (unlike P=S and the cyclopentadienyl ring in 1a), which provides energetically favorable conjugation [torsion angle S2—P1—C10—C15 = −3.8 (4)° in 1vs 35.75 (3)° for the equivalent angle in a selected representative molecule with Fe7 in 1a]. The other related independent torsion angles in 1a are −31. (3), −33.9 (3), −27.0 (3), −28.7 (3), 34.8 (3), 35.7 (3)°, for Fe1–Fe6, respectively.All phosphorus atoms in 1 is the most symmetric with the lowest PSSP torsion [−93.68 (8)°] and shows only a moderate deviation from a right angle. The PSSP torsion angles in 1a  are 6–8° wider than in 1. Notably, ten-membered disulfanes have even wider PSSP torsion angles and the difference between them is smaller, −112.89 (11) and 114.9 (4)°, for 2 and 2a, respectively.It is well recognised that PSSP torsion is a characteristic feature of all disulfanes as a class of organic compounds. The structure of X (X = O or S) can be found in the structures of 1 and 2 , 4.2670 (9), 4.2652 (9) or 4.261 (1) Å (for different independent molecules in 1a) for eight-membered rings, to 4.614 (2) in 2 and 4.604 (1) Å in 2a for the ten-membered rings.The transannular P⋯P distances are very similar within the same ring size and increase, from 4.3331 (17) Å in 1 and 1a was recognised by PLATON  Å. The anisyl substituents may have inhibited this kind of interaction.The strongest intermolecular hydrogen-bonding interaction in 2 are mainly based on the anisyl methoxyl CH3O oxygen atoms O3 and O4 and the P=S sulfur atom S3 as acceptors. Hydrogen-bond donors are the anisyl ortho-hydrogen atoms or methylene hydrogen atoms. Moreover, some C—H..π. interactions may play some role in the system, e.g. C16—H16A⋯ring(C20–C25), see Fig. 5Intermolecular interactions in et al., 2016et al., 2015et al., 2017et al., 2004et al., 1991et al., 1993Bisphosphonothioyl disulfanes represent a rather rare class of compounds  solvent system.Eight- and ten-membered cyclic bisanisylphosphonothioyl disulfanes 2,5-Bis(4-methoxyphenyl)-1,6,3,4,2,5-dioxadithiadiphosphocane 2,5-dithione, 1M.p. 441-443 K.2,5-Bis(4-methoxyphenyl)-1,6,3,4,2,5-dioxadithiadiphosphecane 2,5-dithione, 2Yield: 65%, m.p. 415–417 K.1H NMR (CDCl3): 2.20 , 2.25 , 3.89 , 4.37 , 4.89 , 7.01 , 7.87 .13C NMR: 27.21 , 55.46 (s), 67.08 , 114.03 , 125.41 , 132.89 , 163.09 (s).31P{1H} NMR (CDCl3): 89.19 (3JPP = 4 Hz)18H22O4P2S4: 492.0. Found: 492.9 [M+H]+.MS calculated for C1 was refined as an inversion twin with contribution of the second domain equal to 0.45 (17). This explains the ambiguous Flack parameter and is not surprising since we started from achiral substrates. Structure 2 was refined as a two-component rotational twin with twin law: {2 (Q1–Q3 ca 2e Å3), which are close to sulfur atoms , may stem from conformational flexibility of the ring. Note: the structure of 1 was determined at room temperature (due to a failure of our CryoStream unit) not at 120 K as for 2 but we believe it did not influence the qualitative conclusions drawn from the results.Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989018001068/zp2026sup1.cifCrystal structure: contains datablock(s) 2, global, 1. DOI: 10.1107/S2056989018001068/zp20261sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989018001068/zp20262sup3.hklStructure factors: contains datablock(s) 2. DOI: Click here for additional data file.10.1107/S2056989018001068/zp20261sup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018001068/zp20262sup5.cmlSupporting information file. DOI: 1558043, 719124CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


In the title compound, a phenothiazine moiety is linked to a planar carbazole unit (r.m.s. deviation = 0.029 Å) by a C—C single bond. Their mean planes are inclined to one another by 27.28 (5)°, and the phenothiazine moiety possesses a typical butterfly structure with a fold angle of 27.36 (9)° between the two benzene rings. 29H24N2OS, contains a phenothiazine moiety linked to a planar carbazole unit (r.m.s. deviation = 0.029 Å) by a C—C single bond. The phenothiazine moiety possesses a typical non-planar butterfly structure with a fold angle of 27.36 (9)° between the two benzene rings. The dihedral angle between the mean planes of the carbazole and phenothiazine units is 27.28 (5)°. In the crystal, molecules stack in pairs along the c-axis direction, linked by offset π–π interactions [intercentroid distance = 3.797 (1) Å]. There are C—H⋯π interactions present linking these dimers to form a three-dimensional structure.The title compound, C The fold angle of 27.36 (9)° between the two benzene rings of this moiety compares well with the values reported for similar compounds  Å, interplanar distance = 3.5133 (8) Å, slippage = 1.439 Å, Cg5 is the centroid of the C15–C20 ring; symmetry code: (i) −x + 1, −y, −z + 2]. There are also C–H⋯π interactions present linking these dimers to form a three-dimensional structure -10H-phenothiazine-3-carbaldehyde , 9-ethyl-9H-carbazole-3-boronic acid pinnacol ester , Pd(PPh3)4  and K2CO3  under high vacuum was added a mixture of toluene:water (2:1). The resulting mixture was heated to reflux under an N2 atmosphere for ca 24 h. On completion of the reaction (monitored by TLC), it was quenched by addition of saturated double-distilled H2O and extracted with dichloromethane. The organic phases were collected and washed with brine and dried over anhydrous Na2SO4 and then concentrated. The product was purified by column chromatography on silica gel using ethyl acetate:n-hexane  as eluent, to give the title compound as a pale-yellow crystalline solid (yield 80%). It was characterized by 1H NMR, 13C NMR, IR and ESI–MASS. Brown block-like crystals of the title compound were obtained by slow evaporation at room temperature of a solution in dichloromethane and acetonitrile (1:1 v/v).To a mixture of 7-bromo-10-ethyl-10Uiso(H) = 1.5Ueq(C-methyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989017005540/su5357sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017005540/su5357Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017005540/su5357Isup3.cmlSupporting information file. DOI: 1543611CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


In the crystal, molecules are linked by C—H⋯π interactions, forming chains propagating along the  14H15NO4S, the dihydrothiazine ring adopts a distorted sofa conformation with the S atom displaced from the mean plane through the N and C ring atoms by 0.767 (1) Å. The allyl substituent (C—C=C) is inclined to this mean plane by 78.5 (7)° and the acetate group [C(=O)—O—C] by 66.5 (3)°. In the crystal, molecules are linked by C—H⋯π interactions forming chains propagating along the a-axis direction.In the title compound, C The allyl substituent (C—C=C) is inclined to this mean plane by 78.5 (7)° and the acetate group (O=C—O—C) by 66.5 (3)°. Atom N1 has a planar configuration, the sum of the bond angles being 359.1°.The molecular structure of the title compound, A⋯C5 = 2.47 Å in this fragment [the sum of the van der Waals radii °]. The allyl group is orthogonal to the benzothiazine fragment plane while the terminal double bond is synperiplanar to the N1—C12 bond . The steric repulsion between the allyl substituent and the aromatic cycle (short intramolecular contacts H2⋯C12 = 2.77 Å and H12A⋯C2 = 2.83 Å) results in the elongation of the C1—N1 bond [1.411 (5) Å], compared with the mean value of 1.371 Å  of K2CO3 and the mixture was stirred for 30 min. Allyl bromide  was then added and the mixture was stirred for a further 30 min at 298 K. It was then diluted with cold water and acidified with dilute HCl to pH 4. It was extracted with CH2Cl2 (3 × 10 ml). The organic extracts were combined and the solvent removed by distillation (at reduced pressure at the end). The residue was dissolved in 20 ml of hot methanol and filtered over charcoal. The resulting solution was then placed in a freezer (253 K) for 24 h, after which crystals of the title compound were harvested .The synthesis of the title compound, Uiso = 1.5Ueq(C-methyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989016015978/su5329sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989016015978/su5329Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016015978/su5329Isup3.cmlSupporting information file. DOI: 1508990CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:


II atom in the title complex, [Cd(C10H14N2O)2(C8H4NO2)2(H2O)2], is located on an inversion centre and is coordinated by an N2O4 donor set from pairs of water, 4-cyanobenzoate and N,N-diethylnicotinamide ligands.The Cd 10H14N2O)2(C8H4NO2)2(H2O)2], is centrosymmetric and contains two water molecules, two 4-cyanobenzoate (CB) ligands and two diethylnicotinamide (DENA) ligands. All the ligands are coordinated to the CdII atom in a monodentate mode. The four nearest O atoms around the CdII atom form a slightly distorted square-planar arrangement, with the distorted octahedral coordination sphere being completed by the two pyridine N atoms of the DENA ligands at distances of 2.3336 (13) Å. The dihedral angle between the carboxylate group and the adjacent benzene ring is 8.75 (16)°, while the benzene and pyridine rings are oriented at a dihedral angle of 57.83 (5)°. The water molecules exhibit both intramolecular  and intermolecular  O—H⋯O hydrogen bonds. The latter lead to the formation of supramolecular chains propagating along [110].The mononuclear title cadmium complex, [Cd(C N,N′-diethylnicotinamide (DENA) is an important respiratory stimulant 2(C7H4ClO2)2(H2O)2] 2(H2O)2]  is one form of niacin. A deficiency of this vitamin leads to loss of copper from the body, known as pellagra disease. Pellagra patients show unusually high serum and urinary copper levels  and DENA ligands, namely trans-diaquabis(4-cyanobenzoato-κO)biscadmium, [Cd(CB)2(DENA)2(H2O)2], and report herein its crystal structure.The structure–function–coordination relationships of the arylcarboxylate ion in CdII atom located on an inversion centre, one CB ligand, one DENA ligand as well as one water molecule, all ligands coordinating to the CdII atom in a monodentate mode   of the two symmetry-related monodentate CB anions and water O atoms (O4 and O4i) form a slightly distorted square-planar arrangement around the Cd1 atom, while the slightly distorted octahedral coordination sphere is completed by the two pyridine N atoms (N1 and N1i) of two DENA ligands  Å]; the Cd—N bond length is the longest with 2.3336 (13) Å in the CdO4N2 octahedron. The Cd1 atom lies 0.7558 (1) Å below the planar (O1/O2/C1) carboxylate group. The O—Cd—O and O—Cd—N bond angles range from 87.54 (5) to 92.46 (5)°. In the carboxylate groups, the C—O bonds of the coordinating O atoms [C1—O1 = 1.244 (2) Å and C1—O2 = 1.259 (2) Å] are 0.015 (2) Å longer than those of the non-coordinating ones, indicating delocalized bonding arrangements rather than localized single and double bonds. The dihedral angle between the carboxylate group (O1/O2/C1) and the adjacent benzene (C2–C7) ring is 8.75 (16)°, while the benzene and pyridine (N1/C9–C13) rings are oriented at a dihedral angle of 57.83 (5)°.The two carboxylate O atoms  hydrogen bonds (Table 1S(6) hydrogen-bonding motifs  hydrogen bonds  in H2O (50 ml) and diethylnicotinamide  in H2O (10 ml) with sodium 4-cyanobenzoate  in H2O (100 ml). The mixture was filtered and set aside to crystallize at ambient temperature for several days, giving colourless single crystals.The title compound was prepared by the reaction of CdSO2O) were located in a difference Fourier map and were refined freely. The C-bound H atoms were positioned geometrically with C—H = 0.93, 0.97 and 0.96 Å, for aromatic, methylene and methyl H atoms, respectively, and constrained to ride on their parent atoms, with Uiso(H) = k × Ueq(C), where k = 1.5 for methyl H atoms and k = 1.2 for aromatic and methylene H-atoms.Experimental details including the crystal data, data collection and refinement are summarized in Table 210.1107/S2056989016018247/wm5339sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016018247/wm5339Isup2.hklStructure factors: contains datablock(s) I. DOI: 1517222CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The asymmetric unit of the title compound comprises one cafenstrole ligand molecule and one silver nitrate ion. The coordination bonds between silver and oxygen atoms allow a continuous one-dimensional coordination polymer structure along [001]. The three-dimensional architecture is stabilized by C—H⋯O hydrogen bonds and C—H⋯π interactions N,N-diethyl-3-mesitylsulfonyl-1H-1,2,4-triazole-1-carboxamide), a triazole herbicide, leads to the title coordination polymer, [Ag(NO3)(C16H22N4O3S)]n, whose asymmetric unit comprises one cafenstrole ligand molecule, one AgI atom and one nitrate ion. The AgI atom, with a distorted trigonal–pyramidal environment, is coordinated by one nitrogen atom of a triazole ring, two oxygen atoms of a nitrate ion and one oxygen atom of a neighboring nitrate ion. The coordination bonds between silver and oxygen atoms give rise to a one-dimensional (1D) coordination polymer structure along [001]. The dihedral angle between the planes of the triazole and benzene rings is 87.13 (11)°. In the crystal, the coordination polymer is stabilized by C—H⋯O hydrogen bonds and C—H⋯π interactions, resulting in a three-dimensional architecture.The reaction of silver nitrate and cafenstrole ( L; Kang et al., 2015et al., 2001et al., 2013et al., 2014Recently, we have reported the crystal structure of the ligand cafenstrole  [Ag1—N1 = 2.250 (3) Å] and three oxygen atoms  . The Ag1, O5, N1, and O5i atoms form a slightly distorted triangular basal plane with bond angles O5—Ag1—O5i = 106.52 (5), O5—Ag1—N1 = 118.75 (11) and O5i—Ag1—N1 = 134.63 (11)°. The apex atom, O4, deviates considerably from the normal to the basal plane, as indicated by the O4—Ag1—N1 bond angle of 149.66 (10)°. Other bond angles are 48.93 (10) and 67.18 (10)° for O4—Ag1—O5 and O4—Ag1—O5i, respectively. One oxygen atom of the nitrate ion (O6) is not bound to the AgI ion, whereas the other two oxygen atoms of the nitrate ion (O4 and O5) are bound to the AgI ion. One of the bound O atoms (O5) links neighbouring AgI ion ions, thus forming a 1D polymer along [001]. The triazole plane is rotated about the S1–C10 axis in the opposite direction in comparison with free cafenstrol  −x + 1, −y + 1, z + 1 symmetry, resulting in a 1D chain along [001]  interactions , was determined by Min et al. 2(NO3)(C12H8N2)]·2C12H8N2, has a polymeric chain structure, where nitrate ions show similar coordination bonds compared to those in the title compound, but with AgI ions replaced by with PrIII atoms   in methanol (5 mL). The ligand was purchased from the Dr Ehrenstorfer GmbH Company. Single crystals suitable for X-ray crystallography were obtained by slow evaporation of the solvent at room temperature after one week.The title compound was prepared from a mixed solution of the cafenstrole ligand  in acetone (5 mL) and Ag(NOd(C—H) = 0.98 Å, Uiso(H) = 1.5Ueq(C) for methyl group, d(C—H) = 0.99 Å, Uiso(H) = 1.2Ueq(C) for Csp3—H and d(C—H) = 0.95 Å, Uiso(H) = 1.2Ueq(C) for aromatic C—H.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989016016662/vn2117sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989016016662/vn2117Isup2.hklStructure factors: contains datablock(s) I. DOI: 1510357CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Scientific Reports6: Article number: 28177; 10.1038/srep28177 published online: 07052016; updated: 10102016.In this Article the legend for Figure 5 is incomplete.2+ in cytosol was measured using Fura-2 and Ca2+ in ER was measured using FRET-based probe D1ER cameleon in the basal state and after thapsigargin stimulation (100 nM for 1000 sec). Representative Ca2+ measurements in cytosol and ER (left panel). Quantitative ER Ca2+ levels of the average value prior to thapsigargin stimulation  and the minimum value after addition of thapsigargin (Tg(+)) (right panel). Data are means ± SEM, n = 43 for control, n = 50 for IRS1KO, and n = 40 for IRS2KO β-cells. *P < 0.05, **P < 0.01”.“CaShould read:2+ in cytosol was measured using Fura-2 and Ca2+ in ER was measured using FRET-based probe D1ER cameleon in the basal state and after thapsigargin stimulation (100 nM for 1000 sec). Representative Ca2+ measurements in cytosol and ER (left panel). Quantitative ER Ca2+ levels of the average value prior to thapsigargin stimulation  and the minimum value after addition of thapsigargin (Tg(+)) (right panel). Data are means ± SEM, n = 43 for control, n = 50 for IRS1KO, and n = 40 for IRS2KO β-cells. *P < 0.05, **P < 0.01. (b) Immunoblot of SERCA2 and β-actin in control β-cells, IRS1KO β-cells, and IRS2KO β-cells incubated with vehicle or thapsigargin (100 nM) for 8 h. Data are means ± SEM, n = 4. *P < 0.05”.“(a) Ca


The structures of three bicyclic carbohydrates derivatives containing cyclobutanone or cyclolactame beside the pyranose ring are reported and the conformation and configuration established. R,3R,4R,5R,6S)-4,5-bis(acetyloxy)-7-oxo-2-oxabicyclo[4.2.0]octan-3-yl]methyl acetate, C14H18O8, (I), [-5-acetyloxy-7-hydroxyimino-2-oxobicyclo[4.2.0]octan-4-yl acetate, C11H15NO6, (II), and pyrrol-5-yl]methyl acetate, C14H19NO8, (III), are stable bicyclic carbohydrate derivatives. They can easily be synthesized in a few steps from commercially available glycals. As a result of the ring strain from the four-membered rings in (I) and (II), the conformations of the carbohydrates deviate strongly from the ideal chair form. Compound (II) occurs in the boat form. In the five-membered lactam (III), on the other hand, the carbohydrate adopts an almost ideal chair conformation. As a result of the distortion of the sugar rings, the configurations of the three bicyclic carbohydrate derivatives could not be determined from their NMR coupling constants. From our three crystal structure determinations, we were able to establish for the first time the absolute configurations of all new stereocenters of the carbohydrate rings.The title compounds, [(1 The conformation of the pyranose rings deviates from the ideal chair. The puckering amplitudes and smallest displacement parameters for molecules A and B are q = 0.467 (2)/0.473 (3) Å, θ = 151.9 (4)/150.7 (4)° and φ = 114.1 (6)/114.3 (6)°. The main feature is the absolute configuration of the new stereocenters being 1R and 6S. Surprisingly, the acetyloxy substituents are positioned axially, in contrast to the usual d-gluco arrangement. Obviously the cyclobutanone ring, with its bisectional positioned (C1—C8) and axial bonds (C6—C7) – in relation to the pyranose ring – enforces a flipping of the chair from 4C1 into 1C4. The cyclobutane ring is almost planar [maximum deviation from the best plane of C7 = 0.0762 (15) Å in A and 0.0815 (15) Å in B] and can be described by the dihedral angles, forming by folding along the C6⋯C8 and C1⋯C7 line, between the planes C1–C6–C8/C6–C7–C8  and C1–C6–C7/C1–C7–C8 . The deviation of the carbonyl O atoms (O8A/O8B) from the mean plane of the pyran ring are 0.253 (5) and 0.303 (6) Å in molecules A and B, respectively. The dihedral angles between the pyranose rings and the cyclobutane rings are 61.3 (1) and 62.1 (1)° for molecules A and B, respectively. Four non-classical intramolecular hydrogen bonds for each of the both molecules can be observed A is disordered . Molecules A, B and C mainly differ in the torsion angles C10—O4—C5—C6  and O4—C5—C6—C1  of the acetyloxy substituents in the 5-position /0.763 (3)/0.75 (3) Å, θ = 90.9 (6)/91.0 (2)/91 (2)° and φ = 12.6 (6)/12.7 (2)/28 (3)° for molecules A, B and C, and not the usual chair conformation. This arrangement is caused by the cyclobutane ring with the C1—C8 and C6—C7 bonds, which are bisectional related to the arabinose ring. The absolute configuration on the stereocenters of the shared ring atoms is C1S and C6R. The cyclobutane rings are almost planar with maximum deviations from the best plane of 0.045 (3) Å (C7A), 0.039 (1) Å (C7B) and 0.072 (12) Å (C7C). The nitrogen atoms deviate marginally from these planes [N1A −0.224 (9) Å, N1B 0.199 (4) Å, N1C 0.30 (4) Å. The dihedral angles within the four-membered rings between C1/C6/C8 and C6/C7/C8 are 9.4 (5)° (A), 8.2 (2)°, (B) and 15 (2)° (C), and between C1/C6/C7 and C1/C7/C8 they are 9.0 (5)° (A), 7.9 (3)° (B) and 14 (2)° (C). The hydroxyl group of the oxime substituent can adopt two different configurations. Molecule B exhibits an E configuration. For disordered molecules A and C, the E/Z ratio of the isomers is 0.802 (7):0.198 (7). Thus, the major component (A) is E configured, with the hydroxyl group pointing away from the six-membered ring. In the minor Z isomer (C), the hydroxyl group exhibits a sterically unfavourable interaction with the carbohydrate ring. An intramolecular hydrogen bond between C5A/C5C and O5A is observed on Fig. 2. The pyred Fig. 2.S configured. The six-membered and the five-membered rings are fused in the cis configuration. The C3a—C3 bond is axial and the C7a—N1 bond is bisectionally positioned with respect to the pyranose ring. The pyranose ring exhibits a strongly distorted chair conformation, with puckering parameters q = 0.555 (3) Å, θ = 20.4 (3)° and φ = 267.9 (9)°. The usual d-gluco configuration in the chair form 4C1 is found, in contrast to (I)Compound (III)A molecules are hydrogen-bonded via C4A—H4A⋯O3Ai interactions screwing around the b-axis direction . The A and B molecules are linked by two further hydrogen bonds (C10B—H104⋯O8Aii and C10B—H106⋯O7Aii).The crystal packing of (I)on Fig. 4. BetweenA molecules are in an alternating arrangement with those consisting only of B molecules, both screwing along the b-axis direction on Fig. 5. In contvia weak C—H⋯O hydrogen bonds running along the a-axis direction. The chains formed this way are connected in a pairwise fashion by strong N1—H1A⋯O8 bonds along c et al.  was synthesized from tri-O-acetyl-d-glucal, commercially available or obtained by the procedure of Ferrier  in dry diethyl ether (30 mL) at room temperature over 30 min under an N2 atmosphere. After completion of the reaction (TLC control), zinc dust  was added at 273 K. Acetic acid (13 mL) was added within 10 min and the reaction mixture was stirred for 60 min. The reaction mixture was diluted with diethyl ether (60 mL) and the insoluble materials were filtered off through Celite, which was washed with diethyl ether (5 × 50 mL) and methanol (50 mL). The filtrate was extracted with (3 × 100 mL) water. The organic layer was dried over MgSO4 and concentrated in vacuo. The resulting residue was purified by column chromatography (hexane/ethyl acetate 5:1) to afford pure cyclobutanone (I)rier 1965. TrichlOxime (II) was synthesized from di-O-acetyl-d-arabinal, obtained by the procedure of Ferrier  the corresponding cyclobutanone was synthesized as described above and isolated by column chromatography (hexane/ethyl acetate 5:1) in 83% yield. 242 mg (1.0 mmol) of this cyclobutanone was dissolved in ethanol (2 mL) and then added to a solution of sodium acetate  and hydroxylamine hydrochloride  in water (2 mL). The reaction mixture was stirred at 327 K for 2 h and then for 1 h at room temperature. The reaction mixture was washed with water (30 mL) and extracted with CH2Cl2 (3 × 50 mL). The organic layers were combined, dried over MgSO4, filtered and concentrated in vacuo. The oxime (II)rier 1965. StartinLactam (III) was synthesized from cyclobutanone (I)2Cl2 (3 × 50 mL). The organic layers were combined, dried over MgSO4, filtered and concentrated in vacuo. Thionyl chloride  was added to a solution of the crude oxime in 1,4-dioxane (4 mL), and stirred for 10 min at room temperature. The reaction was quenched with saturated aqueous NaHCO3 (50 mL), and extracted with EtOAc (3 × 100 mL). The organic extracts were washed with brine, dried over MgSO4, and concentrated in vacuo. The residue was purified by column chromatography (hexane/ethyl acetate 1:4) to afford the lactam in analytically pure form . Single crystals suitable for X-ray diffraction were prepared by slow evaporation of a solution of (III)A, caused by flipping of the N—OH group. That disorder also causes disorder of the nearby ring atoms. Therefore the ring atoms of both the five- and six-membered rings were included in the disorder (but the OAc groups were left out). The geometry of the minor component was restrained to be similar to that of the major one with SAME, SADI and SIMU 0.01 restraints. The refinement of the occupation factors revealed an occupation ratio of 0.802 (7)/0.198 (7) for the two disordered components  = 1.2Ueq(C) with the exception of methyl hydrogen atoms, which were placed in their expected positions with HFIX 137 and refined with Uiso(H) = 1.5Ueq(C). For the minor disordered component in compound (II)2 groups (HFIX 13 and 23) and 0.83 Å for OH groups (HFIX 147), and with Uiso(H) = 1.2Ueq(C) and 1.5Ueq(O). Crystal data, data collection and structure refinement details are summarized in Table 4In compound (II)10.1107/S2056989016018727/zl2684sup1.cifCrystal structure: contains datablock(s) I, II, III, global. DOI: 10.1107/S2056989016018727/zl2684Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016018727/zl2684IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989016018727/zl2684IIIsup4.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S2056989016018727/zl2684Isup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016018727/zl2684IIsup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016018727/zl2684IIIsup7.cmlSupporting information file. DOI: 1518715, 1518714, 1518713CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information: