BIOLOGICAL AND BIOCHEMICAL PROPERTIES OF GLYCOSYLATED HUMAN GROWTH HORMONE

我们的长期目标是阐明个体人类生长激素（hGH）变异在健康和疾病状态下人体生理调节中的结构和作用。为了实现这一目标，我们建议研究一种新发现的hGH变体：抗巯基乙醇（MER） 45 kDa hGH的结构和生物学作用。由于它的新颖性，我们对其结构和生物学作用的认识还存在空白。该应用程序的中心假设是，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。所提出的研究背后的基本原理是，最近研究的生长激素和催乳素的结构变异具有与其各自主要亚型不同的生物活性。为了验证这一假设，本研究计划将重点确定MER 45 kDa HGH的结构和生物活性。MER 45 kDa hGH的催乳素样和gh样特性将通过一系列具有代表性的体外和体内生物测定来评估。具体目的是：1)确定MER 45 kDa hGH的结构；2)在一系列测量生长激素作用的生物学分析中确定MER 45 kDa生长激素的生物学效力；3) MER 45 kDa hGH的结构与生物学功能的关系；4)生成抗mer 45 kDa hGH单克隆抗体，用于开发同种异型特异性免疫分析，这将是识别其生理调节的重要工具。在本研究的最后，我们期望能够确定MER 45 kDa hGH的结构，阐明其生理意义，确定其结构-活性关系，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-功能关系的认识空白。这些研究可能会产生新的治疗策略。

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.