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: 6240212
• 负责人: LUIS S HARO
• 金额: $ 10.6万
• 依托单位: UNIVERSITY OF TEXAS SAN ANTONIO
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6240212
• 关键词: carbohydrate metabolism; carbohydrate structure; cell cycle; chemical structure function; electrofocusing; enzyme linked immunosorbent assay; gel electrophoresis; gel filtration chromatography; glucose metabolism; glycosylation; high performance liquid chromatography; ion exchange chromatography; laboratory mouse; laboratory rabbit; laboratory rat; lipid metabolism; lymphatic tissue; molecular cloning; oligosaccharides; protein isoforms; protein sequence; radioimmunoassay; somatotropin; western blottings

This is a proposal requesting support for research and student training to study the biochemistry and biology of glycosylated human growth hormone (G- hGH) which was recently discovered in our laboratory. It is hypothesized that because of its altered structure G-hGH has altered biological activity. To test the hypothesis the hypothesis the research plan will focus on ascertaining the structure and biological activities of G-hGH. The specific objectives are; 1. Structural analysis of G-hGH by : a) micropreparative purification of G-hGH, b) comparative HPLC tryptic peptide mapping of hGH and G-hGH to identify peptide peak shifts that point ot location(s) within hGH that are altered to form G-hGH. c) determination of the amino acid compositions (s) and amino acid sequence(s) of the shifted tryptic peptides, d) identification of hGH glycosylation site(s) by glycopeptide dot blot analysis of shifted tryptic peptide peaks, and e) carbohydrate composition. oligosaccharide mapping and carbohydrate sequence analysis of glycosylated tryptic peptides. 2. Generation of a procedure for preparative isolation of native non-denatured G-hGH using a combination of chromatographic and electrophoretic techniques to provide material for biological testing. 3. Assessment of the potency of G-hGH in a standard hGH radioimmunoassay, a human liver GH radioreceptorassay and in a radioreceptorassay for lactogenic hormones as well as determination of its isoelectric point-molecular weight coordinates in a two-dimensional polyacrylamide gel for its unequivocal identification in the presence of other hGH to promote growth, test its effects on carbohydrate metabolism, measure its influence on fat metabolism,a and assess its ability to stimulate lymphoid cell division. We will also determine the metabolic clearance rate of G-hGH. 5. Develop a radioimmunoassay which is specific for G-hGH with monoclonal antibodies. 6. Research training of graduate and undergraduate students in biomedical research with special emphasis on assisting under-represented minority students to excel in the scientific and biomedical fields. Results of these studies will elucidate the structural nature and physiological significance of G-hGH. This information is important to our current understanding of the structure-function relationships of hGH. Student training will augment the participation of ethnic minorities and women pursuing bio medical careers.

这是一项要求支持研究和学生培训的提案， 研究糖基化人生长激素（G-）的生物化学和生物学， 这是我们实验室最近发现的。 据推测 由于其结构的改变，G-hGH改变了生物学特性， 活动 为了验证假设，研究计划将 重点是确定G-hGH的结构和生物活性。 具体目标是：1。 G-hGH的结构分析：a） G-hGH的微制备纯化，B）比较HPLC胰蛋白酶肽 hGH和G-hGH图谱，以确定肽峰位移， hGH内改变形成G-hGH的位置。 c）决心 的氨基酸组成和氨基酸序列。 转移的胰蛋白酶肽，d）通过 转移的胰蛋白酶肽峰的糖肽斑点印迹分析，和e） 碳水化合物组成 寡糖图谱和糖类 糖基化胰蛋白酶肽的序列分析。 2. 生成 使用制备型分离天然非变性G-hGH的方法 色谱和电泳技术的组合， 生物学试验材料。 3. G-hGH的效价评估 标准hGH放射免疫测定、人肝GH放射受体测定和 催乳激素的放射受体测定以及 它的等电点-分子量坐标在二维 聚丙烯酰胺凝胶，用于在存在 其他hGH促进生长，测试其对碳水化合物代谢的影响， 测量其对脂肪代谢的影响，a并评估其 刺激淋巴细胞分裂。 我们还将确定代谢 G-hGH清除率。 5. 开发一种放射免疫分析， 用单克隆抗体检测G-hGH 6. 研究生科研训练 和生物医学研究的本科生，特别强调 帮助代表性不足的少数民族学生在科学方面取得优异成绩， 和生物医学领域。 这些研究的结果将阐明结构的性质， G-hGH的生理意义 这些信息对我们的 目前对hGH结构-功能关系的理解。 学生培训将增加少数民族的参与， 追求生物医学事业的女性