Assembly and Functions of the PINCH/ILK/CH-ILKBP Complex

PINCH/ILK/CH-ILKBP 复合体的组装和功能

• 批准号: 6741432
• 负责人: CHUANYUE WU
• 金额: $ 22.46万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6741432
• 关键词: actin binding protein; actins; active sites; cell adhesion; cell adhesion molecules; cell differentiation; cell proliferation; enzyme mechanism; enzyme substrate complex; extracellular matrix; extracellular matrix proteins; fibronectins; integrins; molecular assembly /self assembly; phosphotransferases; protein structure function; site directed mutagenesis; tissue /cell culture

DESCRIPTION (provided by applicant): Cell-extracellular matrix adhesion is a fundamental process that regulates cell shape, proliferation and differentiation. Abnormalities in cell-matrix adhesion and extracellular matrix assembly are closely associated with the pathogenesis of a variety of human diseases. The long-term objective of our research is to elucidate the mechanism by which cells regulate cell-matrix adhesion and extracellular matrix assembly. Integrin-linked kinase (ILK) is an important regulator of cell-matrix adhesion and fibronectin matrix assembly. This research project focuses on the molecular mechanism by which ILK functions in these processes. Our hypotheses are that (1) a multi-protein complex comprising ILK, PINCH and CH-ILKBP provides an important physical connection between cell adhesion receptors and the actin cytoskeleton at the cell-matrix contact sites and (2) a PINCH-related protein (PINCH-RP), which was recently cloned by the applicant, regulates the assembly of the PINCH/ILK/CH-ILKBP complex and thereby participates in the regulation of cell adhesion, actin cytoskeleton organization and matrix assembly. The proposed studies are designed to critically test these hypotheses. First, the sites of PINCH, ILK and CH-ILKBP that are involved in the assembly and the localization of the PINCH/ILKICH-ILKBP complex to cell-matrix contact sites will be defined by site-directed mutagenesis. Second, the role of the PINCHIILKICH-ILKBP complex in cell-matrix adhesion, spreading and fibronectin matrix assembly will be determined using reagents that modulate the complex formation. Third, PINCH-RP will be characterized and its potential role in the regulation of the assembly and functions of the PINCHIILKICH-ILKBP complex will be determined. These studies will provide important information on the assembly, function and regulation of the PINCH/ILK/CH-ILKBP complex and will lead to a better understanding of the general mechanism by which cells regulate cell adhesion and matrix assembly, and consequentially, a better understanding of the molecular basis underlying the pathogenesis of diseases associated with abnormal cell adhesion and matrix assembly.

描述（由申请人提供）：细胞-细胞外基质粘附是一种 调节细胞形状、增殖和 分化细胞-基质粘附和细胞外基质 组装与多种人类疾病的发病机制密切相关， 疾病我们研究的长期目标是阐明 细胞通过其调节细胞-基质粘附和细胞外基质组装。 整合素连接激酶（ILK）是细胞-基质粘附的重要调节因子 和纤连蛋白基质组装。该研究项目的重点是分子 ILK在这些过程中发挥作用的机制。我们的假设是 (1)包含ILK、PINCH和CH-ILKBP的多蛋白复合物提供了 细胞粘附受体和肌动蛋白之间的重要物理联系 细胞-基质接触位点的细胞骨架和（2）PINCH相关蛋白 （PINCH-RP），其最近由申请人克隆，调节组装 PINCH/ILK/CH-ILKBP复合物，从而参与调节 细胞粘附、肌动蛋白细胞骨架组织和基质组装。的 拟议的研究旨在严格检验这些假设。一是 参与组装的PINCH、ILK和CH-ILKBP的位点， PINCH/ILKICH-ILKBP复合物定位于细胞-基质接触位点 将通过定点诱变来定义。第二， PINCHILKICH-ILKBP复合物在细胞-基质粘附、铺展和纤连蛋白中的作用 将使用调节复合物的试剂来确定基质组装 阵第三，PINCH-RP将被表征，其潜在的作用， 调节PINCHIILKICH-ILKBP复合物的组装和功能将 被确定。 这些研究将提供有关组装、功能和 PINCH/ILK/CH-ILKBP复合物的调节，并将导致更好的 了解细胞调节细胞粘附的一般机制 和矩阵组装，因此，更好地了解 相关疾病发病机制的分子基础 异常的细胞粘附和基质组装。