Structural Biology of Intramembrane Proteolysis

膜内蛋白水解的结构生物学

• 批准号: 7505302
• 负责人: YIGONG SHI
• 金额: $ 28.18万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7505302
• 关键词: Address; Archaea; Aspartic Endopeptidases; Bacteria; Binding; Biochemical; Caenorhabditis elegans; Classification; Cleaved cell; Complex; Endopeptidases; Escherichia coli; Family; Grant; Homologous Gene; Human; Investigation; Length; Lipid Bilayers; M-protease; Membrane; Metalloproteases; Methionine; Molecular; Organism; Peptide Hydrolases; Peptides; Process; Proteins; Proteolysis; Public Health; Range; Resolution; Roentgen Rays; Serine Protease; Signal Transduction; Signaling Protein; Site; Structure; Work; X ray diffraction analysis; X-Ray Diffraction; improved; inhibitor/antagonist; insight; mutant; presenilin; protease E; rhomboid; signal peptide peptidase; structural biology; three dimensional structure

DESCRIPTION (provided by applicant): Regulated intramembrane proteolysis (RIP) is an ubiquitously conserved signaling mechanism in species ranging from bacteria to humans. An essential step of RIP is the site-specific cleavage of a transmembrane segment in a signaling protein by a specific membrane-embedded protease within the lipid bilayer. These intramembrane proteases are classified into four families: the serine protease rhomboid, metalloprotease Site- 2 protease (S2P), and aspartyl proteases presenilin and signal-peptide peptidase. Despite intense investigation, the structure and mechanisms of these intramembrane proteases remain largely unknown. We have initiated systematic X-ray crystallographic and biochemical analyses of the serine protease rhomboid and the metalloprotease S2P. Significant preliminary results have been achieved; the work proposed here will build on our preliminary results with the following specific aims. (1) Determination of the crystal structure of the transmembrane core domain of the E. coli rhomboid protease GlpG in complex with inhibitor and substrate. (2) Determination of the crystal structure of full-length rhomboid proteases from prokaryotic and eukaryotic species. (3) Determination of the crystal structure of the transmembrane core domain of a S2P intramembrane protease from M. jannaschii. (4) Determination of the crystal structure of the full-length S2P intramembrane protease from M. jannaschii and from E. coli. (5) Determination of the crystal structure of a S2P intramembrane protease in complex with a substrate peptide. These studies, when completed, will reveal significant insights into the structure, function, and mechanism of the rhomboid and S2P families of intramembrane proteases. PUBLIC HEALTH RELEVANCE: Regulated intramembrane proteolysis (RIP) is an ubiquitously conserved signaling mechanism in organisms ranging from bacteria to humans. An essential step of RIP is the site-specific cleavage of a transmembrane segment in a signaling protein by a specific membrane-embedded protease within the lipid bilayer. This proposal seeks to understand the structures and mechanisms of these intramembrane proteases.

描述（由申请人提供）：膜内蛋白水解（RIP）是一种无处不在的信号传导机制，从细菌到人类。 RIP的一个必不可少的步骤是通过脂质双层中特定的膜上包含的蛋白酶在信号蛋白中的跨膜段特异性切割。这些膜内蛋白酶分为四个家族：丝氨酸蛋白酶菱形，金属蛋白酶位点-2蛋白酶（S2P）和aspartyl蛋白酶presenilin和信号肽肽酶。尽管进行了严格的研究，但这些膜内蛋白酶的结构和机制仍然在很大程度上未知。我们启动了丝氨酸蛋白酶菱形和金属蛋白酶S2P的系统X射线晶体学和生化分析。已经取得了重大的初步结果；这里提出的工作将以我们的初步结果为基础，并具有以下特定目标。 （1）确定大肠杆菌菱形蛋白酶GLPG与抑制剂和底物复合物的跨膜核心结构域的晶体结构。 （2）确定原核生物和真核种类全长菱形蛋白酶的晶体结构。 （3）测定Jannaschii的S2P内膜内蛋白酶的跨膜核心结构域的晶体结构。 （4）从M. Jannaschii和大肠杆菌中的全长S2P内膜内蛋白酶的晶体结构测定。 （5）确定与底物肽复合物中膜内蛋白酶的晶体结构。这些研究完成后，将揭示有关膜内蛋白酶的菱形和S2P家族的结构，功能和机制的重大见解。公共卫生相关性：膜内蛋白水解（RIP）是一种无处不在的信号机制，从细菌到人类。 RIP的一个必不可少的步骤是通过脂质双层中特定的膜上包含的蛋白酶在信号蛋白中的跨膜段特异性切割。该建议旨在了解这些膜内蛋白酶的结构和机制。