BIOLOGICAL AND BIOCHEMICAL PROPERTIES OF GLYCOSYLATED HUMAN GROWTH HORMONE

糖基化人类生长激素的生物学和生化特性

• 批准号: 6204140
• 负责人: LUIS S HARO
• 金额: $ 14.87万
• 依托单位: UNIVERSITY OF TEXAS SAN ANTONIO
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6204140
• 关键词: carbohydrate structure; cell cycle; chemical structure function; enzyme linked immunosorbent assay; gel electrophoresis; gel filtration chromatography; glucose metabolism; glycosylation; high performance liquid chromatography; laboratory mouse; laboratory rat; lipid metabolism; lymphatic tissue; monoclonal antibody; oligosaccharides; protein isoforms; protein sequence; radioimmunoassay; somatotropin; western blottings

Our long range goal is to elucidate the structures and understand the roles of individual human growth hormone (hGH) variant sin the regulation of human physiology in health and disease states. As our next objective in pursuit of this goal, we propose in this application to investigate the structure and biological roles of a newly identified hGH variant: mercaptoethanol-resistant (MER) 45 kDa hGH. Due to its novelty, there exists a gap in our knowledge regarding its structure and biological actions. The central hypothesis of the application is that structural components that make MER 45 kDa hGH different from the 22 kDa hGH also make its biological activities different from that of 22 kDa HGH. The rationale behind the proposed research centers on the fact that structural variants of GH and prolactin that have recently been studied possess biological activities that differ from those of their respective major isoforms. To test the hypothesis, this research plan will focus on ascertaining the structure and biological activities of MER 45 kDa HGH. Both the prolactin-like and GH-like properties of MER 45 kDa hGH will be assessed in a set of representative in vitro and in vivo bioassays. The specific objectives are: 1) To determine the structure of MER 45 kDa hGH; 2) To determine the biological potencies of MER 45 kDa hGH in a range of biological assays that are measures of GH action; 3) To correlate the structure of MER 45 kDa hGH to its biological functions; 4) To generate anti-MER 45 kDa hGH monoclonal antibodies for use in development of an isoform-specific immunoassay that will be an important tool in discerning its physiological regulation. At the conclusion of this research, we expect to have determined the structure of MER 45 kDa hGH, elucidate its physiological significance, ascertained it structure-activity relationships and developed an isoform- specific immunoassay The new information gained from the research of the present proposal will fill the gap in our current understanding of the structure-function relationships of MER 45 kDa hGH. New therapeutic strategies could result from these investigations.

我们的长期目标是阐明个体人类生长激素(HGH)变体的结构，并了解其在健康和疾病状态下对人体生理调节中的作用。作为我们追求这一目标的下一个目标，我们建议在这一应用中研究一种新发现的hGH变异体的结构和生物学作用：抗巯基乙醇(MER)45 kDa hGH。由于它的新颖性，我们对它的结构和生物学作用的了解还存在差距。应用的中心假设是，使mer 45 kDa hGH不同于22 kDa hGH的结构成分也使其生物活性不同于22 kDa hGH。这项拟议研究背后的理论基础是，最近研究的生长激素和催乳素的结构变体具有不同于其各自主要异构体的生物活性。为了验证这一假说，本研究计划将重点确定mer 45 kDa HGH的结构和生物活性。45 kDa hGH的催乳素和生长激素样特性将在一系列具有代表性的体外和体内生物检测中进行评估。具体目标是：1)确定mer 45 kDa hGH的结构；2)在一系列衡量GH作用的生物测定中确定mer 45 kDa hGH的生物效价；3)将mer 45 kDa hGH的结构与其生物学功能联系起来；4)制备抗mer 45 kDa hGH的单抗，用于发展一种将成为识别其生理调节的重要工具的异构体特异性免疫分析方法。在本研究的基础上，我们有望确定mer 45 kDa hGH的结构，阐明其生理意义，确定其构效关系，并建立一种异构体特异性免疫分析方法。本研究获得的新信息将填补我们目前对mer 45 kDa hGH结构与功能关系的认识的空白。这些研究可能会产生新的治疗策略。