3-D STRUCTURE DETERMINATION OF SOLUTE TRANSPORTERS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 中心，不一定是研究者的机构。 本计划的目的是通过X射线晶体学研究细菌和真核细胞中的溶质转运机制。我们的项目之一是真核细胞葡萄糖易化转运蛋白的结晶，该项目得到了NIH探索/发展研究资助计划R21的支持。Glut 1是主要易化转运蛋白超家族的成员，是这些易化转运蛋白中被广泛研究的代表。Glut 4负责胰岛素调节的葡萄糖处置。已经鉴定出几种疾病具有导致Gluts功能障碍的突变，例如GLUT 1缺乏综合征和II型糖尿病。GLUT 1已被证明在少数癌细胞中过表达。重要的是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GLUT 1。正在使用高通量机器人方法进行初始结晶。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.