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Vitamin K epoxide reductases (VKORs) are a large family of integral membrane enzymes found from bacteria to humans. Human VKOR, specific target of warfarin, has both the epoxide and quinone reductase activity to maintain the vitamin K cycle. Bacterial VKOR homologs, however, are insensitive to warfarin inhibition and are quinone reductases incapable of epoxide reduction. What affords the epoxide reductase activity in human VKOR remains unknown. Here, we show that a representative bacterial VKOR homolog can be converted to an epoxide reductase that is also inhibitable by warfarin. To generate this new activity, we first substituted several regions surrounding the active site of bacterial VKOR by those from human VKOR based on comparison of their crystal structures. Subsequent systematic substitutions narrowed down to merely eight residues, with the addition of a membrane anchor domain, that are responsible for the epoxide reductase activity. Substitutions corresponding to N80 and Y139 in human VKOR provide strong hydrogen bonding interactions to facilitate the epoxide reduction. The rest of six substitutions increase the size and change the shape of the substrate‐binding pocket, and the membrane anchor domain stabilizes this pocket while allowing certain flexibility for optimal binding of the epoxide substrate. Overall, our study reveals the structural features of the epoxide reductase activity carried out by a subset of VKOR family in the membrane environment. [ABSTRACT FROM AUTHOR]

Background: The mammalian major histocompatibility complex (MHC) harbours clusters of genes associated with the immunological defence of animals against infectious pathogens. At present, no complete MHC physical map is available for any of the wild ruminant species in the world. Results: The high-density physical map is composed of two contigs of 47 overlapping bacterial artificial chromosome (BAC) clones, with an average of 115 Kb for each BAC, covering the entire addax MHC genome. The first contig has 40 overlapping BAC clones covering an approximately 2.9 Mb region of MHC class I, class III, and class IIa, and the second contig has 7 BAC clones covering an approximately 500 Kb genomic region that harbours MHC class IIb. The relative position of each BAC corresponding to the MHC sequence was determined by comparative mapping using PCR screening of the BAC library of 192,000 clones, and the order of BACs was determined by DNA fingerprinting. The overlaps of neighboring BACs were cross-verified by both BAC-end sequencing and co-amplification of identical PCR fragments within the overlapped region, with their identities further confirmed by DNA sequencing. Conclusions: We report here the successful construction of a high-quality physical map for the addax MHC region using BACs and comparative mapping. The addax MHC physical map we constructed showed one gap of approximately 18 Mb formed by an ancient autosomal inversion that divided the MHC class II into IIa and IIb. The autosomal inversion provides compelling evidence that the MHC organizations in all of the ruminant species are relatively conserved. [ABSTRACT FROM AUTHOR]

A novel fluorescent covalent organic framework containing trimethyl silicon group (COF-TMS) was designed and prepared via the Povarov reaction to be used to detect fluorine ions, on account of the interaction between the silicon and the fluorine ions. Trimethyl silicon, as an electron-donating functional group, was replaced with hydrogen atom and the electronic state of the whole material changed. In this process, the change of COF-TMS structure resulted in the fluorescence color changing from red to blue obviously. Furthermore, the structure and optical property of COF-TMS did not change when there are anions such as H2PO4−, ClO4−, SO42−, CO32−, Cl−, Br−, I−, and NO3− in the solution. Notably, the crystallinity also became particularly high in this process, with perfectly regular hexagonal pore structures being clearly observed in the micron range. This work provides a new idea for the future study of COFs in the field of fluorescence through the combination of COFs structure and fluorescence color transformation. [ABSTRACT FROM AUTHOR]

Although many prokaryotes have glycolysis alternatives, it's considered as the only energy-generating glucose catabolic pathway in eukaryotes. Here, we managed to create a hybrid-glycolysis yeast. Subsequently, we identified an inositol pyrophosphatase encoded by OCA5 that could regulate glycolysis and respiration by adjusting 5-diphosphoinositol 1,2,3,4,6-pentakisphosphate (5-InsP 7) levels. 5-InsP 7 levels could regulate the expression of genes involved in glycolysis and respiration, representing a global mechanism that could sense ATP levels and regulate central carbon metabolism. The hybrid-glycolysis yeast did not produce ethanol during growth under excess glucose and could produce 2.68 g/L free fatty acids, which is the highest reported production in shake flask of Saccharomyces cerevisiae. This study demonstrated the significance of hybrid-glycolysis yeast and determined Oca5 as an inositol pyrophosphatase controlling the balance between glycolysis and respiration, which may shed light on the role of inositol pyrophosphates in regulating eukaryotic metabolism. [Display omitted] • Hybrid of glycolysis with the phosphoketolase pathway made an efficient cell factory • Oca5, identified as an inositol pyrophosphatase, degrading 5-InsP 7 to InsP 6 • InsPs control gene expression involved in glycolysis and respiration to balance fluxes • The mechanism of Oca5 regulating glycolysis and respiration was elaborated Glycolysis is considered as the only energy-generating glucose catabolic pathway in eukaryotes. Qin et al. establish a hybrid-glycolysis yeast that disrupts the Embden-Meyerhof-Parnas glycolysis pathway and introduced components of the phosphoketolase pathway. They further identify Oca5 as an inositol pyrophosphatase controlling the balance between glycolysis and respiration activities. [ABSTRACT FROM AUTHOR]

[Display omitted] • The COF-Tr-BA was prepared for selective adsorption and Li-S battery. • COF-Tr-BA shows excellent selectivity for genistein isomer by chemical interaction. • The introduction of boric acid group helps to improve electrochemical performance. Functional triazine-based covalent organic frameworks (COFs) with boric acid units, COF-Tr-BA, are designed and synthesized via cycloaddition of imine-linked COFs, COF-Tr, with (4-ethynylphenyl) boronic acid. Owing to the chemical interaction between boric acid units and adjacent dihydroxyl groups, COF-Tr-BA is used as the adsorbent for the selective adsorption of genistein isomer 4′6,7-trihydroxyisoflavone, showing an excellent selectivity and great adsorption performance with 5.4 times adsorption capacity (67.5 mg g−1) as high as 4′5,7-trihydroxyisoflavone (12.5 mg g−1). Meanwhile, COF-Tr-BA as the sulfur host for lithium-sulfur (Li-S) battery also exhibits a better electrochemical property, giving an initial discharge capacity up to 1349 mA h g−1 and a substantial capacity of 627 mA h g−1 after 200 cycles at 0.5C. It is attributed to the sulfiphilic interaction between the introduced boric acid groups and polysulfides (PSs), as well as the lithiophilic interaction between N atoms from quinoline and triazine moieties on the backbone of COF-Tr-BA and lithium ions (Li+), thus alleviating the dissolution and reducing the shuttle effect of LiPSs. The boric acid functionalized material COF-Tr-BA with the dual-function for selective adsorption and Li-S battery proposed here lays a significant foundation for multifunctional COFs as a versatile platform in practical application. [ABSTRACT FROM AUTHOR]

Human neuroblastoma SH-SY5Y cells are a widely-used human neuronal cell model in the study of neurodegeneration. A recent study shows that, 1-methyl-4-phenylpyridine ion (MPP), which selectively causes dopaminergic neuronal death leading to Parkinson's disease-like symptoms, can reduce SH-SY5Y cell viability by inducing H2O2 generation and subsequent TRPM2 channel activation. MPP-induced cell death is enhanced by increasing the TRPM2 expression. By contrast, increasing the TRPM2 expression has also been reported to support SH-SY5Y cell survival after exposure to H2O2, leading to the suggestion of a protective role for the TRPM2 channel. To clarify the role of reactive oxygen species (ROS)-induced TRPM2 channel activation in SH-SY5Y cells, we generated a stable SH-SY5Y cell line overexpressing the human TRPM2 channel and examined cell death and cell viability after exposure to H2O2 in the wild-type and TRPM2-overexpressing SH-SY5Y cells. Exposure to H2O2 resulted in concentration-dependent cell death and reduction in cell viability in both cell types. TRPM2 overexpression remarkably augmented H2O2-induced cell death and reduction in cell viability. Furthermore, H2O2-induced cell death in both the wild-type and TRPM2-overexpressing cells was prevented by 2-APB, a TRPM2 inhibitor, and also by PJ34 and DPQ, poly(ADP-ribose) polymerase (PARP) inhibitors. Collectively, our results show that increasing the TRPM2 expression renders SH-SY5Y cells to be more susceptible to ROS-induced cell death and reinforce the notion that the TRPM2 channel plays a critical role in conferring ROS-induced cell death. It is anticipated that SH-SY5Y cells can be useful for better understanding the molecular and signaling mechanisms for ROS-induced TRPM2-mediated neurodegeneration in the pathogenesis of neurodegenerative diseases. [ABSTRACT FROM AUTHOR]