CHARACTERIZATION OF GLUCOSE TRANSPORTER FUNCTION

葡萄糖转运蛋白功能的表征

• 批准号: 3225030
• 负责人: CHAN Y. JUNG
• 金额: $ 9.83万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-07-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3225030
• 关键词: affinity labeling; chemical binding; chemical structure function; conformation; crosslink; diabetes mellitus; glucose transport; high performance liquid chromatography; human tissue; insulin sensitivity /resistance; laboratory mouse; laboratory rabbit; mass spectrometry; membrane proteins; molecular pathology; obesity; protein sequence; protein structure function; transport proteins

The long term goal of this proposal is to identify the molecular defect(s) underlying the insulin-resistance seen in human diabetes and obesity. More immediate goals are to understand the molecular mechanisms how the facilitative glucose transporters, a family in intrinsic transmembrane proteins, catalyze translocation of selected sugar molecules. We will use purified human erythrocyte glucose transporter (HEGT) and study its tertiar structure, particularly that of putative glucose channel. A hydropathy analysis of the amino acid sequence of this protein predicts that it contains a transmembrane domain made of twelve transmembrane segments, five of which are amphipathic and may line the purported aqueous channel. It also contains three intracellular and one extracellular nonmembrane domains We use this prediction as a working model in determining the channel structure. We will determine which of these twelve transmembrane segments are positioned in contact with the lipid bilayer, and which the aqueous channel. We will first establish the basic chemical methodology required for the studies. This will include separation and identification of the twelve transmembrane segments. To determine the native tertiary structure of these transmembrane segments. We will chemically crosslink and identify neighboring transmembrane segments, and identify the channel-forming segments and nonchannel-forming segments by measuring the accessibility of segments to transportable, covalently active glucose analogs (4ADG and 6ADG and lipophilic covalent probes, respectively, Based on these data, we will deduce the overall structure of the transmembrane domain. There are strong indication for the importance of intrinsic activity- regulation of glucose transporter function in glucose homeostasis in normal and diseased states. Understanding of this regulation of the intrinsic activity at the molecular level would be greatly assisted once we understan the tertiary structure of the glucose channel as proposed here.

本提案的长期目标是识别分子缺陷 这是人类糖尿病和肥胖症中出现的胰岛素抵抗的基础。 更 当前的目标是了解分子机制， 促进性葡萄糖转运蛋白，一个内源性跨膜转运蛋白家族， 蛋白质，催化选定的糖分子的易位。 我们将使用 纯化的人红细胞葡萄糖转运蛋白（HEGT），并研究其对人红细胞葡萄糖转运蛋白（HEGT） 结构，特别是假定的葡萄糖通道。 水疗法 对这种蛋白质的氨基酸序列的分析预测， 包含由十二个跨膜区段组成的跨膜结构域，五个跨膜区段， 它们是两亲性的并且可以衬在声称的水通道中。 它 也含有三个胞内和一个胞外非膜结构域 我们使用这个预测作为一个工作模型，在确定通道 结构 我们将确定这12个跨膜片段中 被定位成与脂质双层接触， 频道 我们将首先建立所需的基本化学方法 为了研究 这将包括分离和识别 十二个跨膜片段。 为了确定天然三级结构 这些跨膜片段。 我们将化学交联并识别 相邻的跨膜片段，并确定通道形成 段和非渠道形成段，通过测量的可达性 片段转化为可转运的共价活性葡萄糖类似物（4ADG和6ADG 和亲脂性共价探针，分别，基于这些数据，我们将 推断跨膜结构域的整体结构。 有强烈的迹象表明内在活动的重要性- 正常人葡萄糖稳态中葡萄糖转运蛋白功能的调节 和病态的国家 理解这种内在的调节 一旦我们了解了这些分子水平的活动， 葡萄糖通道的三级结构。