Membrane-associated Steps in Coenzyme B12 Biosynthesis

辅酶 B12 生物合成中的膜相关步骤

基本信息

批准号：
8795905
负责人：
Norbert K Tavares
金额：
$ 4.27万
依托单位：
UNIVERSITY OF GEORGIA
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-01-11 至 2015-01-10
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8795905
关键词：
Acids Active Sites Address Anabolism Animals Archaea Area Award Bacteria Binding Biochemistry Bioinformatics Biological Assay C-terminal Calorimetry Cell membrane Cells Chemical Structure Chemicals Chemistry Coenzyme M Coenzymes Collaborations Complex Crystallization Disease Engineering Enzymes Fluorescence Genetic Genetic Complementation Test Goals Human Imagination Ions Kinetics Knowledge Laboratories LacZ Genes Learning Life Location Malignant Neoplasms Membrane Membrane Proteins Metabolism Microscopy Molecular Multiprotein Complexes N-terminal Nobel Prize Pathway interactions Phenotype Phosphoric Monoester Hydrolases Physiological Prokaryotic Cells Property Proteins Reaction Reporter Research Salmonella enterica Scientist Societies Structural Biologist TEV protease Testing Time Titrations Transferase Ultracentrifugation Variant Western Blotting Work antimicrobial base chemical property cobamamide crosslink design fight against improved in vitro Assay in vitro activity in vivo inhibitor/antagonist insight interdisciplinary approach pathogen small molecule

项目摘要

DESCRIPTION (provided by applicant): Coenzyme B12 (aka, adenosylcobalamin, AdoCbl) is synthesized only by prokaryotes, and is essential to the survival of many pathogens. Hence, precise knowledge of the biochemistry underpinning this major pathway (~25 reactions), and an understanding of the structural properties of the enzymes involved, is critical to the design of inhibitors that could be used to target disease-causing bacteria or any other prokaryote of societal importance. I will apply a multidisciplinary approach to study the last step of AdoCbl biosynthesis, which is catalyzed by the AdoCbl-P phosphatase (CobC) enzyme. There are two parts to my proposal: i) Functional and topological studies of CobC; ii) analysis of interactions of CobC with other proteins suspected to comprise a multiprotein complex localized to the cell membrane. I will collaborate with structural biologists led by Ivan Rayment (Department of Biochemistry, UW-Madison) to gain insights into the mechanism of function of CobC, and to advance our understanding of the implications of the existence of a membrane associated biosynthetic complex.

描述（由申请人提供）：辅酶B12（aka，腺苷胆素，adocbl）仅由原核生物合成，对于许多病原体的存活至关重要。因此，对这一主要途径（约25种反应）的生物化学的精确知识以及对所涉及酶的结构特性的理解对于可用于靶向引起疾病细菌的抑制剂或任何其他社会重要性的原核的抑制剂至关重要。我将采用多学科方法来研究ADOCBL生物合成的最后一步，该方法是由AdoCBL-P-P磷酸酶（COBC）酶催化的。我的建议有两个部分：i）COBC的功能和拓扑研究； ii）分析COBC与其他蛋白质的相互作用，该蛋白质涉嫌构成局部在细胞膜上的多蛋白络合物。我将与由Ivan Rayment（UW-Madison生物化学系）领导的结构生物学家合作，以了解COBC功能机理，并促进我们对膜相关生物合成复合物存在的含义的理解。