STRUCTURE OF A MEMBRANE TRANSPORTER WITH INHIBITOR

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 CLC氯转运蛋白协调适当的神经元，肌肉，心血管和上皮功能所必需的氯的运动。 E. coli CLC（ClC-ec 1）与Fab片段复合的抗体（PDB 1 OTS）。 在NIH资助的工作中（1 R 01 GM 070773 - 01 A2），我们发现了CLC-ec 1的抑制剂DIDS，其具有约30 μ M的表观亲和力。 在这项工作中，我们建议确定的ClC-ec 1/DIDS复合物的结构。 这些实验的结果将提供CLC抑制剂结合位点的第一个结构。 这种结构将有助于使用DIDS作为一种工具来探测CLC-ec 1中的氯离子转运机制，并且也可以证明用于绘制DIDS抑制的真核同源物中的DIDS结合位点。 此外，这种结构可能有助于设计更高亲和力的抑制剂，这是CLC非常缺乏的。

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 CLC chloride-transport proteins orchestrate the movement of chloride necessary for proper neuronal, muscular, cardiovascular, and epithelial function. The structure of an E. coli CLC (ClC-ec1) in complex with a Fab fragment has been determined (pdb 1OTS). In work funded by the NIH (1R01GM070773-01A2), we have discovered an inhibitor of CLC-ec1, DIDS, which has an apparent affinity of ~30 uM. In this work, we propose to determine the structure of the ClC-ec1/DIDS complex. The results of these experiments will provide the first structure of a CLC inhibitor binding site. This structure will facilitate the use of DIDS as a tool to probe the chloride-transport mechanism in CLC-ec1, and could also prove useful for mapping the DIDS-binding site in the eukaryotic homologs that are inhibited by DIDS. In addition, the structure may aid in designing higher-affinity inhibitors, which are sorely lacking for the CLCs.