INSULIN RECEPTOR METABOLISM IN 3T3-L1 AND 3T3-C2 CELLS

3T3-L1 和 3T3-C2 细胞中的胰岛素受体代谢

• 批准号: 3228996
• 负责人: BRENT Clyde REED
• 金额: $ 10.29万
• 依托单位: LOUISIANA STATE UNIV HSC SHREVEPORT
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3228996
• 关键词: adipocytes; affinity chromatography; calmodulin; clathrin; crosslink; electrofocusing; fibroblasts; gel electrophoresis; hormone regulation /control mechanism; insulin; insulin inhibitor; insulin receptor; laboratory mouse; laboratory rabbit; membrane activity; phosphates; photochemistry; proteolysis; receptor mediated endocytosis; scintillation counter; sialate; tissue /cell culture; tolbutamide

The reduction of the number of cell-surface insulin receptors by insulin contributes to insulin resistance. A current model for receptor regulation proposes that insulin shifts the distribution of surface receptor to internal sites, increasing receptor degradation in some cells. The hypoglycemic drug tolbutamide counteracts this process by unknown mechanisms increasing the level of surface receptor expressed by the cell. The proposal will continue an evaluation of the steps in receptor processing regulated by insulin and tolbutamide in 3T3-L1 adipocytes. The kinetics of surface receptor movement will be assessed by quantitating both receptor association with clathrin coated vesicles (coated pits) isolated with anticlathrin antibody, and its transit between surface and internal pools represented by its respective sensitivity or resistance to proteolysis. The effects of tolbutamide and insulin on the internal concentration and rate of degradation of labelled receptor will be measured and their relationship established. These studies and further analysis of photolabelled receptor metabolism should define the site of action of these agents and determine the role of receptor occupancy and dissociation in normal receptor regulation. The relationship of receptor location to phosphate and sialic acid content, and to modifications affecting pI and size will be monitored. Protein interacting with the receptor also will be identified by adsorbing labelled cell extracts to purified receptor bound to nitrocellulose or affinity supports, and by in situ crosslinkling of labelled cellular proteins to receptor in intact or permeabilized cells. Proteins interacting with the receptor in an insulin or tolbutamide regulated manner will be purified and further characterized. In both phases receptor will be isolated and quantitated by immunoprecipitation and gel electrophoresis. These studies should more directly define the kinetics and routes of receptor processing, identify the important receptor interactions or modifications involved, and their role in mediating the effects of insulin and tolbutamide on receptor processing. These studies should provide better understanding of the effects of some insulin resistant disease states and their treatment on insulin receptor metabolism and function.

胰岛素减少细胞表面胰岛素受体的数量

项目成果

3T3-L1 adipocyte glucose transporter (HepG2 class): sequence and regulation of protein and mRNA expression by insulin, differentiation, and glucose starvation.

3T3-L1 脂肪细胞葡萄糖转运蛋白（HepG2 类）：胰岛素、分化和葡萄糖饥饿对蛋白质和 mRNA 表达的序列和调节。

• DOI: 10.1016/0003-9861(90)90490-p
• 发表时间: 1990
• 期刊: Archives of biochemistry and biophysics
• 影响因子: 3.9
• 作者: Reed,BC;Shade,D;Alperovich,F;Vang,M
• 通讯作者: Vang,M

An analysis of the relationship between the cellular distribution and the rate of turnover for the separate classes of unoccupied, noncovalently occupied, and covalently occupied insulin receptor.

分析不同类别的未占据、非共价占据和共价占据的胰岛素受体的细胞分布和周转率之间的关系。

• 发表时间: 1989
• 期刊: The Journal of biological chemistry
• 作者: Reed,DK;Newton,C;Fraga,M;Glastad,K;Bagheri,A;Harris,S;Reed,BC
• 通讯作者: Reed,BC