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组分有组织运输重要性的一个显著例子。这些细胞必须与两种不同的环境相互作用，一种在其顶面，另一种在其底面/侧面。Goldenring实验室以前的工作表明，小GTdR ab 11 a和非常规分子马达肌球蛋白Vb的复合物在极化上皮细胞中质膜再循环囊泡的顶端运输中很重要。将利用酵母双杂交筛选补充的特异性和随机突变分析的组合，以确定肌球蛋白Vb与其结合伙伴Rab 11 a和Rab 11-FIP 2之间相互作用的结构基础。此外，内源性肌球蛋白Vb马达复合物的组分将通过从富含H/K-ATP酶的小管囊泡中免疫沉淀马达而从人胃壁细胞中分离，并且复合物的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，这反过来又将促进我们对极化细胞膜再循环的一般机制的理解。