Dynamics of the Vascular Smooth Muscle Cytoskeleton

血管平滑肌细胞骨架的动力学

• 批准号: 7467317
• 负责人: KATHLEEN G MORGAN
• 金额: $ 171.46万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-10 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7467317
• 关键词: Dynamics; Vascular; Smooth; Muscle; Cytoskeleton

DESCRIPTION (provided by applicant) :This is a new application for an interdisciplinary program project led by a cohesive group of investigators. The central theme of the Program Project is to understand the nature and regulation of the actin noncontractile cytoskeletal dynamics in the differentiated vascular smooth muscle cell (dVSMC). Preliminary data already collected by this working group demonstrate that the actin cytoskeleton of the dVSMC is, indeed, dynamic. Additionally, key molecular insights into the function of actin binding proteins in the nonmuscle cytoskeleton, published in part by members of this group, have positioned the field to allow major advances in our understanding of this system over the next five years. This program, in essence, aims to determine the integrative protein biology of the dVSMC cytoskeleton, a goal too broad for an individual RO1, but ideally suited for the PPG mechanism. Project 1 will focus at the level of dVSM tissue and cells, using techniques including protein knockdown, decoy peptides, quantitative imaging, in-cell FRET and crosslinking to investigate the mechanisms of actin remodeling, testing the hypothesis that remodeling of the noncontractile cytoskeleton modulates VSM contractility. Project 2 will focus on physical protein chemical and cell biology approaches to study phosphorylation-induced alterations of the conformation of actin binding proteins and test the hypothesis that conformational changes explain synergistic protein-protein interactions. Project 3 aims to understand the structural and mechanical effects of actin binding proteins (ABP) on the cytoskeleton of dVSMCs using helical reconstruction and single-particle analysis and laser trap methods. Project 4 will test the hypotheses that tandem WH2 domains function in filament nucleation, and that the adjacent proline-rich regions participate in filament elongation and will determine the mechanism by which alpha-actinin and plectin interact with actin using X-ray crystallography, biophysical methods, EM and cell biology. Core B will support all projects by producing custom peptide and protein tools and providing expertise and training in the use of sophisticated biophysical equipment. Lay Summary: The interdisciplinary, integrative, approach of the program will allow individual protein sequences to be directly related to vascular function and, thus, will identify entirely novel targets for the possible development of new therapeutics for the treatment of cardiovascular disease.

描述（由申请人提供）：这是由一个有凝聚力的研究人员团队领导的跨学科计划项目的新申请。该计划项目的中心主题是了解分化血管平滑肌细胞（dVSMC）中肌动蛋白非收缩性细胞骨架动力学的性质和调节。该工作组已经收集的初步数据表明 dVSMC 的肌动蛋白细胞骨架确实是动态的。此外，该小组成员部分发表的关于非肌肉细胞骨架中肌动蛋白结合蛋白功能的关键分子见解，为该领域奠定了基础，使我们在未来五年内对该系统的理解取得重大进展。该计划本质上旨在确定 dVSMC 细胞骨架的整合蛋白质生物学，这个目标对于单个 RO1 来说过于广泛，但非常适合 PPG 机制。项目1将聚焦dVSM组织和细胞水平，利用蛋白敲低、诱饵肽、定量成像、细胞内FRET和交联等技术研究肌动蛋白重塑机制，检验非收缩性细胞骨架重塑调节VSM的假设收缩性。项目 2 将重点关注物理蛋白质化学和细胞生物学方法，以研究磷酸化诱导的肌动蛋白结合蛋白构象的改变，并测试构象变化解释协同蛋白质-蛋白质相互作用的假设。项目 3 旨在利用螺旋重建、单粒子分析和激光陷阱方法了解肌动蛋白结合蛋白 (ABP) 对 dVSMC 细胞骨架的结构和机械效应。项目 4 将测试串联 WH2 结构域在细丝成核中起作用的假设，以及相邻的富含脯氨酸的区域参与细丝伸长，并将使用 X 射线晶体学、生物物理方法确定 α-肌动蛋白和凝集素与肌动蛋白相互作用的机制、 EM 和细胞生物学。 Core B 将通过生产定制肽和蛋白质工具并提供使用复杂生物物理设备的专业知识和培训来支持所有项目。 概要：该项目的跨学科、综合性方法将使个体蛋白质序列与血管功能直接相关，从而为可能开发治疗心血管疾病的新疗法确定全新的靶点。