PROTEOGLYCAN MEDIATED SIGNALING IN CELL ADHESION

细胞粘附中蛋白聚糖介导的信号传导

• 批准号: 6138476
• 负责人: JOHN R. COUCHMAN
• 金额: $ 20.89万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-01-01 至 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6138476
• 关键词: CHO cells; G protein; active sites; affinity chromatography; affinity labeling; biological signal transduction; cell adhesion; cell migration; cytoskeleton; fibronectins; glycation; integrins; intermolecular interaction; lipid metabolism; membrane lipids; membrane proteins; phosphatidylinositols; phosphorylation; protein kinase C; protein structure function; protein tyrosine kinase; proteoglycan; recombinant proteins; syndecan; transfection

DESCRIPTION: Anchorage-dependent cells respond to matrix molecules by the formation of focal adhesions. These are sites of complex transmembrane signaling which impact cell behavior and growth, both through short term effects on cytoskeletal organization and longer term effects on gene transcription. The long term goal of this research is to understand the molecular mechanisms underlying focal adhesion formation and subsequent signaling role in migration, anchorage, and matrix assembly and turnover. Cell-matrix interactions have important roles in development and wound repair, and are altered in the pathogenesis of rheumatic diseases, tumorigenesis and fibrosis. Syndecan-4 is a transmembrane heparan sulfate proteoglycan that regulates focal adhesion assembly. Transfection studies show that a region of its cytoplasmic domain can influence focal adhesions, cytoskeletal organization, and migration. This region of syndecan-4, but not equivalent regions from the other three syndecan members, can directly bind protein kinase C and regulate its activity. Oligomerization of the core protein is required. This, and protein kinase C regulatory ability, are both augmented by syndecan-4 interactions with phosphatidylinositol 4,5 biphosphate (PIP2), known to be a regulator of actin filament and focal adhesion assembly. The proposed studies will determine: (1) whether protein kinase C, syndecan-4 and PIP2 form a ternary signaling complex in vivo, (2) the sites of interaction between these three cell membrane associated components and how this is regulated, (3) the roles of the ectodomain and cytoplasmic domain of syndecan-4 core proteins, and of its glycanation, in focal adhesion assembly, and (4) the pathway by which protein kinase C regulates focal adhesion assembly, possibly through convergence with other signaling cascades, such as G proteins of the Ras superfamily, lipid metabolism and tyrosine kinases. These studies will use a combination of physical, chemical, immunological and molecular techniques, and will serve as a basis for intervention in the controlling role of focal adhesion and cytoskeletal organization.

描述：贴壁依赖性细胞通过以下方式对基质分子做出反应： 粘着斑的形成。 这些是复杂跨膜的位点 通过短期影响细胞行为和生长的信号传导 对细胞骨架组织的影响和对基因的长期影响 转录。 这项研究的长期目标是了解 粘着斑形成及其后续的分子机制 在迁移、锚定、基质组装和周转中的信号作用。 细胞-基质相互作用在发育和伤口中发挥重要作用 修复并改变风湿性疾病的发病机制， 肿瘤发生和纤维化。 Syndecan-4 是一种跨膜硫酸乙酰肝素 调节粘着斑组装的蛋白多糖。 转染研究 表明其细胞质结构域的一个区域可以影响粘着斑， 细胞骨架组织和迁移。 Syndecan-4 的这个区域，但是 与其他三个多聚糖成员不等效的区域，可以直接 结合蛋白激酶 C 并调节其活性。 寡聚化 需要核心蛋白。 这，和蛋白激酶C的调节能力， 两者均通过 syndecan-4 与磷脂酰肌醇 4,5 的相互作用得到增强 二磷酸盐 (PIP2)，已知是肌动蛋白丝和焦点的调节剂 粘合组装。 拟议的研究将确定：（1）是否 蛋白激酶 C、syndecan-4 和 PIP2 形成三元信号复合物 体内，（2）这三个细胞膜之间相互作用的位点 相关组件及其监管方式，（3） syndecan-4核心蛋白及其胞外域和胞质域 聚糖化，在粘着斑组装中，以及（4）途径 蛋白激酶 C 可能通过调节粘着斑组装 与其他信号级联的融合，例如 Ras 的 G 蛋白 超家族、脂质代谢和酪氨酸激酶。 这些研究将使用 物理、化学、免疫学和分子技术的结合， 并将作为干预重点控制作用的基础 粘附和细胞骨架组织。