Molecular Genetics of Intracellular Protein Transfer

细胞内蛋白质转移的分子遗传学

• 批准号: 8126455
• 负责人: Chris Alan Kaiser
• 金额: $ 45.34万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-02-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8126455
• 关键词: Address; Biochemical; Biochemistry; Cells; Chemistry; Coupled; Cysteine; Diagnosis; Disulfides; Electron Transport; Electron Transport Pathway; Endoplasmic Reticulum; Enzymes; Eukaryotic Cell; Extracellular Protein; Feedback; Gene Proteins; Genetic; Glutathione; Health; Homologous Gene; Investigation; Link; Mammalian Cell; Membrane; Membrane Proteins; Molecular; Molecular Genetics; Organism; Oxidants; Oxidases; Oxidation-Reduction; Pathway interactions; Physiological; Protein Complex Subunit; Protein Disulfide Isomerase; Proteins; Reactive Oxygen Species; Regulation; Saccharomyces cerevisiae; Succinate Dehydrogenase; Sulfhydryl Compounds; Work; Yeasts; disulfide bond; insight; novel; oxidation; parallel processing; polypeptide; protein folding; protein structure; research study; secretory protein; small molecule

DESCRIPTION (provided by applicant): The endoplasmic reticulum (ER) is the compartment where membrane and secretory proteins are modified, folded, and assembled. A key step in the folding of most extracellular proteins includes formation of disulfide bonds, which can add stability to folded polypeptides and can link together subunits of protein complexes. Previous work on this project has delineated the core pathway for protein disulfide bond formation in eukaryotic cells in which disulfide bonds generated by an oxidase, Ero1p, are then transferred to protein disulfide isomerase (PDI), which in turn transfers a disulfide bond to substrate proteins. Through a combination of biochemical, structural, and genetic experiments in the yeast S. cerevisiae we have gained fundamental insight into the molecular mechanisms that underlie each of the steps in this pathway. In this application we now focus on understanding how the protein oxidation pathway is integrated into the redox biochemistry of the cell. Aims of the proposal include: understanding how Pdi1p and related proteins regulate Ero1p activity, investigation of a novel block in ER translocation instigated by hyperactive Ero1p activity, and elucidation of a new electron transport chain in the ER coupled to disulfide bon formation. PUBLIC HEALTH RELEVANCE: In S. cerevisiae it will be possible to apply the full power of a well developed genetic organism to uncover the genes and proteins responsible protein folding in the ER. A detailed understanding of these pathways in S. cerevisiae will make it possible to understand parallel processes in mammalian cells, opening the way to diagnose dysfunctional folding in the ER of mammalian cells and possible detrimental effects of reactive oxygen species generated by the disulfide bond formation pathway.

描述（由申请人提供）：内质网（ER）是膜和分泌蛋白修饰、折叠和组装的隔室。大多数细胞外蛋白质折叠的关键步骤包括二硫键的形成，二硫键可以增加折叠多肽的稳定性，并可以将蛋白质复合物的亚基连接在一起。该项目的先前工作已经描绘了真核细胞中蛋白质二硫键形成的核心途径，其中由氧化酶Ero1p产生的二硫键然后转移到蛋白质二硫键异构酶（PDI），后者又将二硫键转移到底物蛋白质。通过对酵母S.通过对酿酒酵母的研究，我们对该途径中每个步骤的分子机制有了基本的了解。在这个应用程序中，我们现在专注于了解蛋白质氧化途径是如何整合到细胞的氧化还原生物化学。该提案的目的包括：了解Pdi1p和相关蛋白质如何调节Ero1p活性，研究一种新的阻断ER易位煽动过度Ero1p活性，并阐明一种新的电子传递链在ER耦合到二硫键形成。公共卫生相关性：在S。通过研究酿酒酵母，将有可能应用发育良好的遗传生物的全部力量来揭示ER中负责蛋白质折叠的基因和蛋白质。对S.酿酒酵母的研究将使人们有可能了解哺乳动物细胞中的平行过程，开辟了诊断哺乳动物细胞ER功能失调折叠和二硫键形成途径产生的活性氧可能产生的有害影响的途径。