STRUCTURE OF A MEMBRANE TRANSPORTER WITH INHIBITOR

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 研究中心，而研究中心不一定是研究者所在的机构。 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. 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 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.