Myosin Vb involvement in plasma membrane recycling

肌球蛋白 Vb 参与质膜回收

• 批准号: 6998756
• 负责人: JOSEPH Thomas Edward ROLAND
• 金额: $ 4.4万
• 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/6998756
• 关键词: cellular polarity; enzyme mechanism; gastrointestinal epithelium; guanosinetriphosphatases; human tissue; hydrogen potassium exchanging ATPase; immunoprecipitation; intracellular transport; mass spectrometry; membrane activity; myosins; postdoctoral investigator; protein protein interaction; protein structure function; yeast two hybrid system

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：极化上皮细胞是细胞成分有组织运输重要性的一个生动例子。这些细胞必须与两种不同的环境相互作用，一种在其顶端表面，一种在其基底/侧面。 Goldenring 实验室之前的工作表明，小 GTPase Rab11a 和非常规分子运动肌球蛋白 Vb 的复合物对于极化上皮细胞中质膜回收囊泡的顶端运输非常重要。将结合特异性和随机突变分析并辅以酵母双杂交筛选，以确定肌球蛋白 Vb 与其结合伴侣 Rab11a 和 Rab11-FIP2 之间相互作用的结构基础。另外，内源性肌球蛋白Vb运动复合物的组分将从人胃壁细胞中通过对富含H/K-ATP酶的管状囊泡的运动进行免疫沉淀来分离，并且将通过质谱确定复合物成员的身份。这项工作的结果将有助于发现肌球蛋白 Vb 的调节机制，从而促进我们对极化细胞膜回收的一般机制的理解。