MOLECULAR BIOLOGY & FUNCTION OF PLATELET MEMBRANE GP130

在二维SDS凝胶上可见的40种或更多蛋白质中， 血小板膜蛋白，只有少数有名字， 分配的功能。 大多数目前的特征 糖蛋白，包括膜糖蛋白（GP）Ib和 GPIIb-IIIa复合物，在特定的 病理状态，这一特征极大地有助于描述 其结构和功能。 最近出现的重组 然而，DNA技术使研究许多 其他以前没有特征的和潜在的非常重要的 膜蛋白 我们使用这些技术来调查 血小板共有的膜糖蛋白， 内皮细胞，并获得了一些有趣的 互补DNA（cDNA）克隆。 本研究的目的是完全描述一种新的 描述了膜糖蛋白，称为GP 130，我们有 在人血小板和脐静脉内皮细胞中鉴定。 分子生物学、生物化学和 免疫学技术将用于提供详细的 了解这个分子的结构和功能。 到 为了完成这些目标，将进行以下研究 进行。 首先，全长的完整核苷酸序列 将确定编码GP 130的cDNA克隆。 这些克隆 将允许糖蛋白的氨基酸序列 推导 第二，cDNA克隆将用作杂交 探针来表征大小，染色体组织， 染色体位置和编码GP 130的基因数量。 最后，GP 130在血小板功能中的作用将是 研究了 多克隆和单克隆抗体， GP 130将被生产和使用，以深入了解精确的 这种膜糖蛋白在细胞中的亚细胞定位和功能 人体血小板 GP 130可能与其他 膜成分或与下面的细胞骨架将 也可以探索。 生物化学性质，包括有限的氨基 酸序列数据，以及糖基化的程度， GP 130将被分析。 这三个相互关联的项目共同构成了一个研究 该计划应该产生有价值的，及时的新信息， 新描述的血小板的详细结构和功能 膜糖蛋白 重要的是，这项调查是 预计将作为研究其他重要问题的原型， 存在于血小板和其他细胞中的膜抗原 不能用更经典的生物化学方法来研究 技术. 这样的研究将进一步加深我们对 血小板膜结构和功能，这反过来又应该 导致输血治疗、血小板储存和 我们对血小板功能的理解和管理 紊乱

Of the 40 or more proteins visible on a two-dimensional SDS gel of platelet membrane proteins, only a handful have names and assigned functions. Most of the currently characterized glycoproteins, including membrane glycoprotein (GP) Ib and the GPIIb-IIIa complex, are missing or defective in a particular pathological state, a feature that has greatly aided in delineating their structure and function. The recent advent of recombinant DNA technology, however, has made it possible to study many other previously uncharacterized and potentially very important membrane proteins. We have used these techniques to investigate membrane glycoproteins that are shared by platelets and endothelial cells, and have obtained a number of interesting complementary DNA (cDNA) clones. The purpose of the study is to characterize completely a newly described membrane glycoprotein, termed GP130, that we have identified in human platelets and umbilical vein endothelial cells. A combination of molecular biological, biochemical, and immunological techniques will be used to provide a detailed understanding of the structure and function of this molecule. To complete these objectives, the following studies will be conducted. First, the complete nucleotide sequence of full-length cDNA clones that encode GP130 will be determined. These clones will permit the amino acid sequence of the glycoprotein to be deduced. Second, the cDNA clones will be used as hybridization probes to characterize the size, chromosomal organization, chromosomal location, and number of genes that encode GP130. Finally, the role of GP130 in platelet function will be investigated. Polyclonal and monoclonal antibodies specific for GP130 will be produced and used to gain insight into the precise subcellular location and function of this membrane glycoprotein in human platelets. The possible association of GP130 with other membrane components or with the underlying cytoskeleton will also be explored. Biochemical properties, including limited amino acid sequence data, as well as the degree of glycosylation of GP130, will be analyzed. Together, these three interrelated projects constitute a research program that should yield valuable, timely new information about the detailed structure and function of a newly described platelet membrane glycoprotein. Importantly, this investigation is expected to serve as as prototype for the study of other important membrane antigens present in platelets and other cells that have not been amenable to study by more classical biochemical techniques. Studies such as these will further our knowledge of platelet membrane architecture and function, which should in turn lead to improvements in transfusion therapy, platelet storage, and our understanding and management of platelet functional disorders.