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Structural biology of biotin-dependent carboxylases

生物素依赖性羧化酶的结构生物学

基本信息

批准号：
8008942
负责人：
VIVIEN YEE
金额：
$ 6.2万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-01-15 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8008942
关键词：
Acidity Active Sites Address Affect Amino Acid Substitution Amino Acids Animal Welfare Bacteria Bibliography Binding Biochemical Biological Models Biotin Carbon Dioxide Carboxyltransferases Catabolism Catalysis Catalytic Domain Cobalt Complex Country Critiques Crystallization Data Decarboxylation Dependence Disease Environment Environmental Impact Enzymes Equipment Fatty Acids Genes Gluconeogenesis Human IACUC Image In Vitro International Ions Label Lead Ligand Binding Ligands Metabolic Metabolic Diseases Metabolic Pathway Methods Methylmalonyl-CoA carboxyltransferase Missense Mutation Monitor Multienzyme Complexes Mutation N1&apos -carboxybiotin Nature Outcome Oxaloacetates Patients Play Population Preparation Principal Investigator Propionic Acids Proteins Protons Publishing Pyruvate Pyruvates Raman Spectrum Analysis Reaction Relative (related person)Research Research Design Research Ethics Committees Resolution Resources Role Sampling Shapes Site-Directed Mutagenesis Spectrum Analysis Staining method Stains Stretching Structure Techniques Testing Vertebrates Yeasts abstracting biotin 2 carbon dioxide transport carboxylation cofactor deprotonation design developmental disease enolate enzyme deficiency expiration human subject inhibitor/antagonist ketotic hyperglycinemia lactic acidemia methylmalonyl-CoA decarboxylase novel oxidation particle programs propionyl-coenzyme A protein folding research study response structural biology tiglyl-coenzyme A tool

项目摘要

Biotin-dependent carboxylases use a covalently attached biotin cofactor to transport carbon dioxide as carboxybiotin. Four human biotin-dependent carboxylases have been identified, and play central roles in metabolic pathways such as oxidation of odd-chain fatty acids, catabolism of branched amino acids, fatty acid synthesis, and gluconeogenesis. The bacterial transcarboxylase enzyme has long served as a model system for the human biotindependent carboxylases. This proposal focuses on structure-function studies of transcarboxylase and on two human enzymes, propionyl coenzyme A carboxylase and methylcrotonoyl coenzyme A carboxylase. The broad objective is to provide structural information for these enzymes in order to better understand their assembly as multi-enzyme complexes and their mechanisms of catalytic activity. The specific aims are to: 1. To probe the chemistry and mechanism of the transcarboxylase active sites with structural studies of the isolated 12S and 5S subunits. Crystals of the isolated subunits bound to substrate, product, or cofactor will be studied by X-ray crystallography and Raman crystallography. Site- directed mutagenesis will be used to probe the importance of residues highlighted by structural studies. 2. To probe the molecular architecture of the intact transcarboxylase multienzyme complex by X-ray crystallography and electron microscopy. 3. To investigate the crystal structure of human propionyl coenzyme A carboxylase and to model the possible structural consequences of deficiency mutations in the metabolic disorder propionic acidemia. 4. To investigate the crystal structure of the human methylcrotonyl coenzyme A carboxylase beta subunit and to model the possible structural consequences of deficiency patients with the metabolic disorder 3-methylcrotonylglycinuria. Relevance: Biotin-dependent enzymes are important in human metabolism. Mutations which alter their genes are found in patients with metabolic and developmental disorders. Investigating the structures of these enzymes will aid understanding of how these proteins assemble and function, and help explain how mutations may cause disease.

生物素依赖性羧化酶使用共价附着的生物素辅助因子作为羧基生物素运输二氧化碳。四、人类生物素依赖性