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Haloalkane dehalogenases (HLDs) are enzymes that catalyze the cleavage of carbon-halogen bonds by a hydrolytic mechanism. Although comparative biochemical analyses have been published, no classification system has been proposed for HLDs, to date, that reconciles their phylogenetic and functional relationships. In the study presented here, we have analyzed all sequences and structures of genuine HLDs and their homologs detectable by database searches. Phylogenetic analyses revealed that the HLD family can be divided into three subfamilies denoted HLD-I, HLD-II, and HLD-III, of which HLD-I and HLD-III are predicted to be sister-groups. A mismatch between the HLD protein tree and the tree of species, as well as the presence of more than one HLD gene in a few genomes, suggest that horizontal gene transfers, and perhaps also multiple gene duplications and losses have been involved in the evolution of this family. Most of the biochemically characterized HLDs are found in the HLD-II subfamily. The dehalogenating activity of two members of the newly identified HLD-III subfamily has only recently been confirmed, in a study motivated by this phylogenetic analysis. A novel type of the catalytic pentad (Asp-His-Asp+Asn-Trp) was predicted for members of the HLD-III subfamily. Calculation of the evolutionary rates and lineage-specific innovations revealed a common conserved core as well as a set of residues that characterizes each HLD subfamily. The N-terminal part of the cap domain is one of the most variable regions within the whole family as well as within individual subfamilies, and serves as a preferential site for the location of relatively long insertions. The highest variability of discrete sites was observed among residues that are structural components of the access channels. Mutations at these sites modify the anatomy of the channels, which are important for the exchange of ligands between the buried active site and the bulk solvent, thus creating a structural basis for the molecular evolution of new substrate specificities. Our analysis sheds light on the evolutionary history of HLDs and provides a structural framework for designing enzymes with new specificities. 相似文献
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JoHsi Huang Hannah KC Co YiChen Lee ChiaChou Wu Shenghong Chen 《Molecular systems biology》2021,17(10)
Cells metabolize nutrients through a complex metabolic and signaling network that governs redox homeostasis. At the core of this, redox regulatory network is a mutually inhibitory relationship between reduced glutathione and reactive oxygen species (ROS)—two opposing metabolites that are linked to upstream nutrient metabolic pathways (glucose, cysteine, and glutamine) and downstream feedback loops of signaling pathways (calcium and NADPH oxidase). We developed a nutrient‐redox model of human cells to understand system‐level properties of this network. Combining in silico modeling and ROS measurements in individual cells, we show that ROS dynamics follow a switch‐like, all‐or‐none response upon glucose deprivation at a threshold that is approximately two orders of magnitude lower than its physiological concentration. We also confirm that this ROS switch can be irreversible and exhibits hysteresis, a hallmark of bistability. Our findings evidence that bistability modulates redox homeostasis in human cells and provide a general framework for quantitative investigations of redox regulation in humans. 相似文献
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Ling Tong Seong Heon Kim Kristin Rosner Wensheng Yu Bandarpalle B. Shankar Lei Chen Dansu Li Chaoyang Dai Vinay Girijavallabhan Janeta Popovici-Muller Liping Yang Guowei Zhou Aneta Kosinski M. Arshad Siddiqui Neng-Yang Shih Zhuyan Guo Peter Orth Shiying Chen Joseph A. Kozlowski 《Bioorganic & medicinal chemistry letters》2017,27(14):3037-3042
We have identified a series of hydantoin-derived TNF-a converting enzyme (TACE) inhibitors containing a pendant fused bi-heteroaryl group, which demonstrate sub-nanomolar potency (Ki), excellent activity in human whole blood assay, and improved DMPK profiles over prior series. 相似文献
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K S Kosinski L Molthan L White 《Revue fran?aise de transfusion et immuno-hématologie》1984,27(5):619-624
We present three examples of anti-Fy3 produced by Negroes. Results of serologic studies were similar to those of previously reported examples of Negro produced anti-Fy3, including negative to weak reactivity with cord red blood cells, separable anti-Fya, and the presence of multiple unexpected alloantibodies. These findings reinforce the difference between anti-Fy3 produced by Negroes and that produced by non-Negroes. The rare formation of Duffy antibodies by Negroes may signify the presence of an amorphic gene at the Duffy locus. 相似文献
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