Myosin Vb involvement in plasma membrane recycling

肌球蛋白 Vb 参与质膜回收

• 批准号: 7325683
• 负责人: JOSEPH Thomas Edward ROLAND
• 金额: $ 4.99万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-12-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7325683
• 关键词: Apical; Back; Binding; Cell membrane; Cells; Complement; Complex; Endocytosis; Environment; Epithelial Cells; Family; Gastric Parietal Cells; H(+)-K(+)-Exchanging ATPase; Human; Ion Channel; Ion Pumps; Lateral; Mass Spectrum Analysis; Membrane; Molecular Motors; Monomeric GTP-Binding Proteins; Motor; Myosin ATPase; Play; Protein Array; Proteins; Recycling; Regulation; Role; Screening procedure; Surface; System; Vesicle; Work; Yeasts; base; member; polarized cell; rab11 protein; receptor; trafficking; yeast two hybrid system

Polarized epithelial cells are a dramatic example of the importance of the organized transport of cellular components. These cells must interact with two distinct environments, one on their apical surface and one on their basal/lateral surfaces. Previous work in the Goldenring lab has shown that the complex of the small GTPase Rab11a and the unconventional molecular motor myosin Vb are important in the apical transport of plasma membrane recycling vesicles in polarized epithelial cells. A combination of specific and random mutational analysis complemented by yeast two-hybrid screening will be utilized in order to determine the structural basis of the interaction between myosin Vb and its binding partners Rab11a and Rab11-FIP2. Additionaly, the components of the endogenous myosin Vb motor complex will be isolated from human gastric parietal cells by immunoprecipitaion of the motor from H/K-ATPase-enriched tubulovesicles, and the identities of the members of the complex will be determined by mass spectroscopy. Results from this work will aid in discovering the mechanisms of regulation of myosin Vb, which in turn will advance our understanding of the general mechanisms of membrane recycling in polarized cells.

极化上皮细胞是细胞有组织运输重要性的一个生动例子。 成分。这些细胞必须与两种不同的环境相互作用，一种位于其顶端表面，另一种位于其顶端表面。 在其基部/侧面。 Goldenring 实验室之前的工作表明，小分子的复杂性 GTPase Rab11a 和非常规分子运动肌球蛋白 Vb 在顶端转运中发挥着重要作用 极化上皮细胞中的质膜回收囊泡。特定与随机的结合 将利用突变分析辅以酵母双杂交筛选来确定 肌球蛋白 Vb 与其结合伙伴 Rab11a 和 Rab11-FIP2 之间相互作用的结构基础。 此外，内源性肌球蛋白 Vb 运动复合体的成分将从人体中分离出来。 通过对富含 H/K-ATP 酶的管泡的运动进行免疫沉淀来检测胃壁细胞，以及 复合体成员的身份将通过质谱法确定。这项工作的结果 将有助于发现肌球蛋白 Vb 的调节机制，从而推动我们的研究 了解极化细胞中膜回收的一般机制。