Conformational changes in CIC chloride transporters

CIC 氯化物转运蛋白的构象变化

• 批准号: 9358552
• 负责人: Joseph A Mindell
• 金额: $ 16.94万
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9358552
• 关键词: Area; Biochemical; Biological Assay; Biological Process; CLC Gene; Chloride Channels; Chloride Ion; Chlorides; Collaborations; Coupled; Crystallography; Electrophysiology (science); Family; Family member; Goals; Homologous Gene; Individual; Ion Channel; Liposomes; Membrane Potentials; Methods; Molecular Conformation; Neurons; Physiological; Property; Proteins; Protons; Rest; Skeletal Muscle; Structure; Subgroup; Universities; antiport; antiporter; improved; pH Homeostasis; proteoliposomes; single molecule

This project continued studying conformational changes in ClC-type chloride channel proteins. The ClC family of chloride-conducting ion channels is involved in a host of biological processes; these channels maintain the resting membrane potential in skeletal muscle, modulate excitability in central neurons, and are involved in the homeostasis of pH in a variety of intracellular compartments. Despite their physiological importance, the mechanisms by which these channels function are poorly understood. We are attempting to understand the functional properties of these proteins by examining several family members, including both eukaryotic and prokaryotic homologs. Our current focus in this area is to observe the flux due to a single ClC transporter in a single proteoliposome. We are doing this using single molecule methods in collaboration with Dimitrios Stamou at the University of Copenhagen. Toward this end we have developed a robust bulk assay of chloride-coupled proton transport, and then subjected individual liposomes to similar conditions. In this circumstances, we observe robust acidification in single liposomes. We are currently optimizing these assays and trying to improve their robustness.

该项目继续研究ClC型氯离子通道蛋白的构象变化。ClC家族的氯离子传导离子通道参与了一系列的生物过程，这些通道维持骨骼肌的静息膜电位，调节中枢神经元的兴奋性，并参与多种细胞内隔室的pH动态平衡。尽管它们具有重要的生理意义，但人们对这些通道的作用机制知之甚少。我们试图通过研究几个家族成员来了解这些蛋白质的功能特性，包括真核和原核同源物。我们目前在这一领域的重点是观察单个蛋白脂质体中单个CLC转运体的通量。我们正在与哥本哈根大学的迪米特里奥斯·斯塔莫合作，使用单分子方法来完成这项工作。为此，我们开发了一种强大的氯化物偶联质子转运的批量分析方法，然后将各个脂质体置于类似的条件下。在这种情况下，我们观察到单一脂质体中的强酸化。我们目前正在优化这些检测，并试图提高它们的稳健性。