权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

BIOCHEMICAL STUDIES ON THE LH-HCG RECEPTOR

LH-HCG 受体的生化研究

基本信息

批准号：
3318768
负责人：
Brij Saxena
金额：
$ 10.65万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
1988
资助国家：
美国
起止时间：
1988-02-01 至 1991-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3318768
关键词：
Primates affinity chromatography chorionic gonadotropin corpus luteum disulfide bond gel electrophoresis genetic manipulation hormone receptor laboratory mouse luteinizing hormone membrane reconstitution /synthesis molecular cloning monoclonal antibody phospholipids protein reconstitution protein sequence protein structure ultracentrifugation

项目摘要

A receptor preparation specific to luteinizing hormone and to human chorionic gonadotropin has been isolated from bovine corpora lutea and is available in our laboratory in sufficient quantities to permit biochemical, structural and immunological studies proposed here. Subunits of the receptor of 85,000 and 38,000 molecular weight are also available in the purified form. The isolated receptor and its subunits are largely protein in nature with minor moieties of carbohydrates and lipids. We propose to perform recombination experiments to confirm that the 85K and 38K fragments are disulfide-linked subunits of the receptor molecule. Reconstitution experiments, where receptors are taken out of the biological membrane and put back into an artificial membrane environment will also be performed for the LH-hCG receptor using phospholipids. We will characterize the protein moieties by SDS-gel electrophoresis, ultracentrifugal analyses, peptide mapping, end group analyses and partial amino acid sequence determination. A library of monoclonal antibodies to the LH-hCG receptor and its subunits will be produced. The antibodies will be purified and mapped for their specificities. The monoclonal antibodies will be utilized in structural analyses of the receptor; in the immunocytochemical localization of the receptor; to follow the biochemical fate of the hormone-receptor complex in the cell. The monoclonal antibodies will be immobolized on a solid phase and used to characterize receptor fractions by affinity chromatography. The monoclonal antibodies will also be used in the cloning of genetic material. The cloned DNA sequences will be utilized to alternatively confirm the sequence of the receptor and subunits. The monoclonal antibodies can be used for the development of more specific assays of the receptor. Monoclonal antibodies, because of their selectivity, specificity and unlimited productibility, can lend themselves for clinical use when coupled to a drug. These studies are vital to understand the mechanism of hormone action at the molecular level during normal and abnormal gonadal function.

一种针对黄体生成素和TO的受体制剂从牛中分离出人绒毛膜促性腺激素黄体库，并在我们的实验室提供足够的允许生化、结构和免疫学的数量在这里提出的研究。受体亚基85,000和纯化形式也可获得38,000个相对分子质量。分离的受体及其亚基本质上主要是蛋白质。含有少量的碳水化合物和脂肪。我们建议进行重组实验以确认85K和 38K片段是受体的二硫键连接的亚基分子。重建实验，其中受体被从生物膜中取出，放回人工的黄体生成素-人绒毛膜促性腺激素也将在膜环境下进行。使用磷脂的受体。我们将对这种蛋白质进行表征部分经十二烷基硫酸钠-凝胶电泳法、超速离心法分析，肽图谱、端基分析和部分氨基酸序列确定。抗肿瘤单抗的文库将产生黄体生成素-人绒毛膜促性腺激素受体及其亚单位。这个抗体将被提纯并绘制其特异性图谱。这个将利用单抗进行结构分析受体的免疫细胞化学定位；追随激素受体复合体的生化命运在牢房里。这些单抗将被固定在固相，用于通过亲和力表征受体组分层析法。这些单抗也将用于遗传物质的克隆。克隆的DNA序列将用来替代地确认受体的序列和子单位。所述单抗可用于治疗开发更具特异性的受体检测方法。单克隆抗体，因为它们的选择性、特异性和无限可生产性，当结合时可用于临床一种药物。这些研究对于了解心力衰竭的发病机制至关重要。正常和正常的激素在分子水平的作用性腺功能异常。