ROLE OF CELL ADHESION IN BIOLOGICAL SIGNAL TRANSDUCTION

细胞粘附在生物信号转导中的作用

• 批准号: 5210031
• 负责人: GALEN B SCHNEIDER
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5210031
• 关键词: affinity chromatography; biological signal transduction; cell adhesion; cell differentiation; dental implants; extracellular matrix; fibroblasts; immunofluorescence technique; immunologic assay /test; immunoprecipitation; osteoblasts; protein purification; protein tyrosine phosphatase; tissue /cell culture; western blottings; wound healing

Focal adhesions (FA) are prominent transmembrane structures of many cells in culture. Individual FA's serve to link the extracellular matrix (ECM) to the actin cytoskeleton, and to initiate a cascade of signalling pathways through integrin receptors. The regulation of these cellular signalling pathways is important with respect to many cellular functions including growth and differentiation, tumorigenesis, wound healing, osseointegration, and the formation of a mineralized matrix. The objective of this study was to evaluate protein tyrosine phosphatase (PTP) activity that may regulate tyrosine phosphorylation in FA's. Using a PTP assay, we have confirmed the observations of Maher (PNAS 90:11177, 1993) that lysates of fibroblasts in suspension have elevated PTP activity when compared with lysates of adherent fibroblasts. Looking for PTP's that may be adhesion regulated, we have identified the cytoplamic PTP, PTPDl, in immunoprecipitates of the FA protein paxillin. We are currently exploring this interaction and the relationship of this PTP to FA's. Microinjection of the cytoplasmic domain of a receptor-type PTP, PTP , (GST fusion protein subclone generously provided by Mathew Thomas, Wash. Univ., St. Louis) into adherent fibroblasts depleted the tyrosine phosphorylation in FA's. However, the FA structure appeared to be stable as judged by staining with anti-vinculin. Our results suggest that the tyrosine phoshorylation in FAs is regulated not only by kinases but also by PTP's.

局灶性粘连（FA）是突出的跨膜粘连 培养中的许多细胞的结构。 每个足协的发球局都把 细胞外基质（ECM）的肌动蛋白细胞骨架，并启动 通过整合素受体的信号传导途径级联。 调控 这些细胞信号通路的重要性，对于许多 细胞功能，包括生长和分化，肿瘤发生， 伤口愈合、骨整合和矿化的形成 矩阵 本研究的目的是评估蛋白酪氨酸 磷酸酶（PTP）活性，可调节酪氨酸磷酸化， FA的。 使用PTP分析，我们证实了Maher的观察结果 (PNAS 90：11177，1993），悬浮液中的成纤维细胞的裂解物具有 与粘附成纤维细胞的裂解物相比，PTP活性升高。 寻找可能受粘附调节的PTP，我们已经确定了 细胞质PTP，PTPD 1，在FA蛋白桩蛋白的免疫沉淀物中。 我们目前正在探索这种相互作用和这种关系， PTP到FA。 显微注射一种受体型的胞质结构域 PTP，PTP，（GST融合蛋白亚克隆，由Mathew慷慨提供 华盛顿州托马斯 大学、St. Louis）转化为粘附的成纤维细胞， FA中的酪氨酸磷酸化。 然而，FA结构似乎 通过抗黏着斑蛋白染色判断稳定。 我们的研究结果表明 FA中的酪氨酸磷酸化不仅受激酶的调节， 也是PTP的。