Characterization of a novel endosomal sorting complex

新型内体分选复合物的表征

• 批准号: 6998565
• 负责人: ERIC DAVID SPEAR
• 金额: $ 4.4万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6998565
• 关键词: Golgi apparatus; SDS polyacrylamide gel electrophoresis; Saccharomyces cerevisiae; aminoacid; cell free system; enzyme activity; enzyme complex; fungal genetics; guanine nucleotide binding protein; intracellular transport; membrane proteins; nutrition; permease; postdoctoral investigator; protein localization; protein transport; vesicle /vacuole

DESCRIPTION (provided by applicant): The regulated sorting of plasma membrane proteins is controlled by extracellular signals. A number of membrane proteins are retained intracellularly until their transport to the plasma membrane is stimulated by a particular signal. The transport of the S. cerevisiae general amino acid permease Gaplp is regulated in this manner. Under nutrient-limiting conditions, Gaplp is transported to the plasma membrane where it has high activity. Conversely, growth in nutrient-rich conditions results in little Gaplp at the plasma membrane, and low permease activity. Recently, a novel complex containing two small GTP-binding proteins has been shown to be required for Gaplp plasma membrane localization and activity (Gao and Kaiser, manuscript in preparation). My aims are to: (1) Determine the intrinsic and activated guanine nucleotide binding and enzymatic properties of the GTPases, (2) Identify additional components of this Gaplp-sorting complex using biochemical and genomic means, and (3) Reconstitute the endosome-to-Golgi transport of Gaplp in vitro using purified membrane fractions and components, with the goal of identifying the requirements for this process.

描述（由申请人提供）：质膜蛋白的调节排序是由细胞外信号控制的。许多膜蛋白保留在细胞内，直到它们被特定信号刺激转运到质膜。酿酒酵母通用氨基酸通透酶Gaplp的转运以这种方式调节。在营养限制条件下，Gaplp 被转运至具有高活性的质膜。相反，在营养丰富的条件下生长会导致质膜上的 Gaplp 很少，并且通透酶活性较低。最近，一种含有两个小 GTP 结合蛋白的新型复合物已被证明是 Gaplp 质膜定位和活性所必需的（Gao 和 Kaiser，手稿正在准备中）。我的目-使用纯化的膜组分和成分进行 Gaplp 的体外高尔基运输，目的是确定该过程的要求。