Structural Studies of Sugar Transporters

糖转运蛋白的结构研究

基本信息

批准号：
8035627
负责人：
DANENG WANG
金额：
$ 9.97万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2011-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8035627
关键词：
Affinity Amino Acids Arginine Binding Binding Sites Biochemical Biochemistry Biological Assay C-terminal Cells Charge Complex Crystallization Data Electron Spin Resonance Spectroscopy Escherichia coli Family Funding Glucose Transporter Glycogen Storage Disease High temperature of physical object Homologous Gene Human Inorganic Phosphate Transporter Kinetics Maps Measures Mediating Membrane Transport Proteins Methods Molecular Conformation Muscle Mutagenesis Mutate Mutation Non-Insulin-Dependent Diabetes Mellitus Organophosphates Pathogenesis Phase Play Process Research Personnel Resolution Role Side Site Spin Labels Structure Substrate Specificity Suspension substance Suspensions Techniques Temperature Testing Transmembrane Transport Vesicle alpha-glycerophosphoric acid conformer glucose 6-phosphate(transporter)glucose uptake improved member novel programs reconstitution structural biology sugar

项目摘要

Membrane transport, for example glucose uptake, Is essential to the cell. Such processes are often mediated by secondary membrane transporters. The largest secondary transporter family is the major facilitator superfamily (MFS), with more than 5,000 members identified to date, including the glucose transporter from muscle (Glut4) and the glucose-6-phosphate transporter (G6PT), both from human and both are involved in the pathogenesis of type 2 diabetes. We propose to study the transport mechanisms of a bacterial homolog of Glut4 and G6PT, the glycerol-3-phosphate transporter (GIpT) from E. coli, using structural biology, biochemistry and spectroscopic approaches. In the previous funding cycle, we determined the crystal structure of GIpT in a substrate-free form at 3.3 A resolution. The structure suggests a "rocker-switch" mechanism for substrate transport. In the next funding cycle, to better understand GIpT's substrate specificity and substrate-induced conformational change, we plan to determine the crystal structures of the transporter in two substrate-bound forms. We will also test the rocker-switch mechanism and examine the role that key amino acid residues play using mutagenesis, transport assays and spectroscopic techniques. The structural and mechanistic information gained from GIpT can be used directly to improve our understanding of Glut4 and G6PT.

膜转运，例如葡萄糖吸收，对细胞至关重要。这样的过程经常被调解由次级膜转运蛋白。最大的二级转运蛋白家族是主要的促进者超家族（MFS），迄今为止有5,000多名成员，包括肌肉（GLUT4）和葡萄糖-6-磷酸转运蛋白（G6PT），都参与 2型糖尿病的发病机理。我们建议研究细菌同源物的运输机制 GLUT4和G6PT，使用结构生物学，来自大肠杆菌的3-磷酸甘油转运蛋白（GIPT）生物化学和光谱方法。在上一个资金周期中，我们确定了晶体 GIPT以3.3 A分辨率为3.3的无基质形式的结构。该结构暗示了一个“摇杆开关” 底物运输机制。在下一个资金周期中，以更好地了解Gipt的基板特异性和底物引起的构象变化，我们计划确定以两种底物结合形式的转运蛋白。我们还将测试摇杆开关机制，并检查关键氨基酸残基使用诱变，传输测定和光谱技术发挥作用的作用。从GIPT获得的结构和机械信息可直接用于改善我们的了解GLUT4和G6PT。