CRYSTAL STRUCTURE DETERMINATION OF YEAST GET3

酵母 GET3 的晶体结构测定

• 批准号: 8170267
• 负责人: BINGDONG SHA
• 金额: $ 0.07万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170267
• 关键词: ATP phosphohydrolase; ATP-Binding Cassette Transporters; Adopted; Binding; C-terminal; Complex; Computer Retrieval of Information on Scientific Projects Database; Family; Funding; Golgi Apparatus; Grant; Institution; Integral Membrane Protein; Ligand Binding; Mediating; Membrane; Molecular Conformation; Nucleotides; Pathway interactions; Proteins; Research; Research Personnel; Resources; Signal Recognition Particle; Source; Structure; Tail; Time; Transmembrane Domain; United States National Institutes of Health; Yeasts; trafficking

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Tail-anchored (TA) proteins represent a unique family of transmembrane proteins which contain a single transmembrane helix (about 25 residues) at the C-terminus. The mechanisms how the TA proteins insert the TMD into membranes are distinct from the well-studied co-translational insertion pathway, which is mediated by the cytosolic signal recognition particle (SRP). The post-translational insertion of the TA proteins into ER membrane requires the Golgi ER trafficking (GET) complex which contains Get1, Get2 and Get3. Get3 is an ATPase and can recognize and bind the C- terminal transmembrane domain (TMD) of the TA proteins. It is not well understood how GET complex accomplish the membrane insertion of the TA protein. Recently we have determined the yeast Get3 crystal structure in the open conformation. The structure suggested that Get3 may adopt an open conformation in nucleotide-free state and a closed conformation in ATP-bound state. The conformational changes to switch the Get3 between the open and closed conformations may facilitate the membrane insertions for TA proteins. Get3 may share a similar mechanism as the ABC transporters to transfer bound ligands. Now we are requesting beam time to determine Get3 crystal structure in the closed conformation to confirm our hypothesis.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 尾锚定 (TA) 蛋白代表一个独特的跨膜蛋白家族，其 C 末端含有单个跨膜螺旋（约 25 个残基）。 TA 蛋白将 TMD 插入膜的机制与经过充分研究的共翻译插入途径不同，后者是由胞质信号识别颗粒 (SRP) 介导的。 TA 蛋白翻译后插入 ER 膜需要高尔基体 ER 运输 (GET) 复合体，其中包含 Get1、Get2 和 Get3。 Get3 是一种 ATP 酶，可以识别并结合 TA 蛋白的 C 端跨膜结构域 (TMD)。目前尚不清楚 GET 复合物如何完成 TA 蛋白的膜插入。最近我们确定了酵母Get3开放构象的晶体结构。该结构表明Get3在无核苷酸状态下可能采用开放构象，而在ATP结合状态下可能采用闭合构象。 Get3 在开放和闭合构象之间切换的构象变化可能有利于 TA 蛋白的膜插入。 Get3 可能与 ABC 转运蛋白具有相似的机制来转移结合的配体。现在我们要求光束时间来确定闭合构象的 Get3 晶体结构，以证实我们的假设。