MOLECULAR BIOLOGY & FUNCTION OF PLATELET MEMBRANE GP130

在二维十二烷基硫酸钠凝胶上可见的40种或更多蛋白质中 在血小板膜蛋白中，只有少数有名字和 分配的职能。大多数目前被描述为 糖蛋白，包括膜糖蛋白(GP)Ib和 GPIIb-IIIa复合体，在特定情况下缺失或有缺陷 病理状态，这一特征极大地有助于描绘 它们的结构和功能。重组技术的最新进展 然而，DNA技术已经使研究许多 其他以前没有特征的，可能非常重要的 膜蛋白。我们已经使用这些技术来调查 血小板膜糖蛋白和 内皮细胞，并获得了一些有趣的 互补DNA(CDNA)克隆。 这项研究的目的是完全描述一种新的 描述了我们所拥有的膜糖蛋白gp130 在人的血小板和脐静脉内皮细胞中被鉴定。 分子生物学、生物化学和 免疫学技术将用于提供详细的 了解这种分子的结构和功能。至 完成这些目标后，将进行以下研究 指挥。第一，全长的核苷酸序列 将确定编码gp130的cDNA克隆。这些克隆人 将允许糖蛋白的氨基酸序列是 推断出来的。第二，克隆的cdna将用于杂交。 用探针来表征大小，染色体组织， 染色体位置和编码gp130的基因数量。 最后，gp130在血小板功能中的作用将是 调查过了。特异性的多克隆和单克隆抗体 Gp130将被制作并用于深入了解 该膜糖蛋白的亚细胞定位和功能 人类的血小板。Gp130与其他基因的可能联系 膜成分或与下面的细胞骨架会 也是被探索的。生化特性，包括有限的氨基酸 酸序列数据，以及糖基化程度 Gp130，将被分析。 这三个相互关联的项目共同构成了一项研究 计划应提供有关以下方面的有价值的、及时的新信息 一种新的血小板的详细结构和功能 膜糖蛋白。重要的是，这次调查是 期望作为其他重要研究的原型 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来研究 技巧。这样的研究将进一步加深我们对 血小板膜的结构和功能，依次 导致输血治疗、血小板储存和 我们对血小板功能的认识和处理 精神错乱。

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.