3-D STRUCTURE DETERMINATION OF SOLUTE TRANSPORTERS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 本研究的目的是用X射线结晶学方法研究细菌和真核细胞中的溶质转运机制。我们的一个项目是真核促进葡萄糖转运蛋白的结晶，该项目得到了美国国立卫生研究院探索/发展研究资助计划R21的支持。Glut1是主要促进剂超家族的成员之一，是这些易化转运蛋白的一个被广泛研究的代表。GLUT4负责胰岛素调节的葡萄糖处置。已发现几种疾病具有导致谷氨酸功能障碍的突变，如GLUT1缺乏综合症和II型糖尿病。Glut1已被证明在少数癌细胞中过表达。重要的是要获得高分辨率的结构，以及在原子水平上表征传输机制。到目前为止，还没有可用于任何葡萄糖促进剂的三维晶体结构。我们直接从人红细胞膜中分离出GLUT1。使用高通量机器人方法的初步结晶正在进行中。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The aim of this proposal is to study solute transport mechanisms in bacterial and eukaryotic cells by x-ray crystallography. One of our projects is the crystallization of eukaryotic facilitated glucose transporter, which was supported by NIH Exploratory/Developmental Research Grant Program R21. Glut1, a member of the Major Facilitator Superfamily, is an extensively studied representative of these facilitated transporters. Glut4 is responsible for insulin-regulated glucose disposal. Several diseases have been identified with mutations resulting in malfunction of Gluts, such as GLUT1 deficiency syndrome and type II diabetes. GLUT1 has been shown to overexpress in few cancer cells. It is important to obtain a high-resolution structure, as well as to characterize the transport mechanism at an atomic level. Thus far, there is no 3-D crystal structure available for any glucose facilitator. We directly isolate GLUT1 from human Red Blood Cells membranes. Initial crystallization using a high throughput robotic approach is being carried out.