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）：调节膜内蛋白水解（RIP）是从细菌到人类等物种中普遍存在的保守信号传导机制。 RIP 的一个重要步骤是通过脂质双层内的特定膜嵌入蛋白酶对信号蛋白中的跨膜片段进行位点特异性切割。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Site-2蛋白酶(S2P)以及天冬氨酰蛋白酶早老素和信号肽肽酶。尽管进行了大量研究，这些膜内蛋白酶的结构和机制仍然很大程度上未知。我们已经开始对丝氨酸蛋白酶菱形和金属蛋白酶 S2P 进行系统的 X 射线晶体学和生化分析。已取得显着的初步成果；这里提议的工作将以我们的初步成果为基础，并实现以下具体目标。 (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)原核和真核物种全长菱形蛋白酶晶体结构的测定。 (3) 测定来自M. jannaschii 的S2P膜内蛋白酶的跨膜核心结构域的晶体结构。 (4) 测定来自 M. jannaschii 和来自 E. coli 的全长 S2P 膜内蛋白酶的晶体结构。 (5)确定与底物肽复合的S2P膜内蛋白酶的晶体结构。这些研究完成后，将揭示对菱形和 S2P 膜内蛋白酶家族的结构、功能和机制的重要见解。公共健康相关性：调节膜内蛋白水解 (RIP) 是从细菌到人类等生物体中普遍存在的保守信号机制。 RIP 的一个重要步骤是通过脂质双层内的特定膜嵌入蛋白酶对信号蛋白中的跨膜片段进行位点特异性切割。该提案旨在了解这些膜内蛋白酶的结构和机制。