Molecular Biological Approach to Nephritogenic Proteins

肾炎蛋白的分子生物学方法

基本信息

批准号：
02807034
负责人：
HIRANO Hideyasu
金额：
$ 1.09万
依托单位：
University of Occupational and Environmental Health, School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至无数据
项目状态：
已结题

项目摘要

Fibronectin is an extracellular adhesive protein which actively functions during wound healing, cell migration, phagocytosis of Kupffer's cell or macrophages, etc. Cellular fibronectin promotes the differentiation of the neural crest cells to adrenergic phenotype of cells. Whereas the myotube formation process induced by the fusion of nyoblast was inhibited by fibronectin. This activity indicated that fibronectin is a differentiation inhibition substance. In the case of chondrocyte cell culture, fibronectin causes dedifferentiation of chondrocytes to a phenotype resembling the nesenchynal cell. Thus the fibronectin has inconsistent functions in the molecule. We thought from these published results that fibronectin could be a controlling morphology protein rather than a differentiation or dedifferentiation substance. Our target was to clone controlling morphology proteins, then we had to exclude many metabolic enzymes distributed in the organs(kidney, liver, lung etc.). We investigated … More to obtain or to discover proteins which change morphology, acting outside or beneath the membrane, through nephritis and fibrosis. We used antibodies against kidney basement membranes to obtain clones carrying cell surface vicinal proteins. Several inserts cDNAs obtained by library screenings of lambda-gtll were used as probes for Northern blot hybridization. The probe purified from 21st clone(lambda NH21)showed an increased level of expression through wound-healing/fibrosis. Sequenced NH21 was homologous to human profilin cDNA. Our nobel approach decreased the obtaining possibility of cytoplasmic metabolic enzyme eDNAs, and at the same time, this gave us to identity controlling morphology proteins speedily and to find out a protein links to signal transduction, that is, one of the obtained clones carried profilin cDNA.The approach using pentapeptide(VIO)which is a part of type III repeat of fibronectin, VIO induced thymocyte differentiation.We have not yet succeeded by the approach to obtain cDNA coding Nephritogenoside using PCR technique and direct hybridization of synthetic oligos to lambda-gtllcDNA library. Less

纤连蛋白是一种细胞外粘附蛋白，在伤口愈合，细胞迁移，kupffer细胞或巨噬细胞的吞噬作用等过程中起作用。细胞纤维蛋白会促进神经元细胞对细胞肾上腺素型的分化。而纤连蛋白抑制了由Nyoblast融合引起的肌管形成过程。该活性表明纤连蛋白是一种分化抑制物质。在软骨细胞培养物的情况下，纤连蛋白会导致软骨细胞的去分化为类似于Nesenchynal细胞的表型。纤连蛋白在分子中具有不一致的功能。我们认为，从这些发表的结果中，纤连蛋白可能是控制形态蛋白的一种，而不是分化或去分化物质。我们的目标是克隆控制形态蛋白，然后我们必须排除分布在器官（肾脏，肝，肺等）中的许多代谢酶。我们研究了……更多地获得或发现通过肾炎和纤维化改变形态，作用在膜外或下方的蛋白质。我们使用针对肾脏基底膜的抗体获得携带细胞表面替代蛋白的克隆。通过Lambda-GTLL的库筛选获得的几个插入物cDNA用作北印迹杂交的探针。从第21个克隆（Lambda NH21）纯化该探针通过伤口愈合/纤维化显示表达水平升高。测序的NH21与人类profilin cDNA同源。我们的诺贝尔方法降低了细胞质代谢酶EDNA的获得潜力，与此同时，这使我们迅速控制了身份，可以迅速控制形态蛋白质，并找出与信号转导的蛋白质联系，即使用p肽肽（vio）III（vio）III，这是一个cdna。胸腺细胞的分化。我们尚未通过使用PCR技术获得cDNA编码肾op的方法，并直接杂交合成寡聚与lambda-gtllcdna库。较少的