Degradation of Mislocalized Secretory and Membrane Proteins

错误定位的分泌蛋白和膜蛋白的降解

• 批准号: 7734851
• 负责人: Ramanujan S Hegde
• 金额: $ 12.24万
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7734851
• 关键词: Affinity Chromatography; Degradation Pathway; Elements; Fractionation; Goals; Homeostasis; In Vitro; Membrane Proteins; Microsomes; Pathway interactions; Preparation; Proteins; Reaction; Risk; System; Ubiquitination; reconstitution

The selective recognition and degradation of mislocalized secretory and membrane proteins has been reconstituted in vitro. In this system, secretory and membrane proteins are synthesized in their non-translocated state simply by omitting ER-derived rough microsomes from the reaction or, if they are included, by inhibiting translocation using cotransin. With either of these manipulations, non-translocated proteins are rapidly ubiquitinated in preparation for their degradation. We are employing both classical fractionation and affinity purification approaches to identify the machinery for the selective recognition and degradation of these non-translocated proteins.

在体外重新构成了错误定位的分泌和膜蛋白的选择性识别和降解。在该系统中，仅通过从反应中省略ER衍生的粗糙微粒体或通过使用cotransin抑制易位，将分泌蛋白和膜蛋白在其非转移状态中合成。使用这两种操纵，非转移的蛋白质被迅速泛素化，以准备降解。我们正在采用经典分级和亲和力纯化方法来识别这些非翻译蛋白的选择性识别和降解的机械。