COBRE: UNR: MOLECULAR PHYSIOL & REGULATION OF VOLUME SENSITIVE CHLORIDE CHANNEL

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 科布雷项目1将研究心脏中容量调节阴离子通道的分子生理学和调节。氯离子通道调节的分子机制对于理解细胞如何调节其体积和了解离子通道蛋白的结构/功能关系是一个重要的问题。我们先前提出ClC-3是CLC电压依赖性氯离子通道超家族的成员，它是负责心肌和平滑肌细胞天然容量调节的外向整流性阴离子通道(VSOAC)的候选分子。本项目中提出的实验将提供新的见解，通过氨基端氨基酸的各种蛋白激酶在调节两种主要的ClC-3亚型中的作用，使用转基因小鼠来进一步研究ClC-3与天然VSOAC的关系并揭示它们的生理作用，最后将使用分子和蛋白质组学方法来鉴定构成天然VSOAC多聚体蛋白复合体的主要成分和辅助蛋白。由于心脏Cl-通道的激活对动作电位时程和自律性产生显著影响，是细胞容量稳态的关键调节因素，因此这些通道在心律失常、心肌缺血、充血性心力衰竭和肥厚等心肌疾病中具有重要的临床意义。该项目对于阐明容量敏感的氯通道在心脏和心血管系统中的正常生理和可能的病理生理学作用具有重要的潜力。

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. COBRE Project 1 will study the molecular physiology and regulation of volume-regulated anion channels in the heart. The molecular mechanism of Cl- channel regulation is an important issue for understanding how cells regulate their volume and for understanding structure/function relationships of ion channel proteins in general. We previously proposed ClC-3, a member of the ClC superfamily of voltage-dependent Cl- channels, as a molecular candidate responsible for native volume-regulated outwardly rectifying anion channels (VSOACs) in cardiac and smooth muscle cells. The experiments proposed in this project will provide new insights into the role of phosphorylation by various protein kinases of amino terminus amino acids in the regulation of the two major ClC-3 isoforms, use transgenic mice to further examine the relationship between ClC-3 and native VSOACs and reveal their physiological role, and finally will use molecular and proteomic approaches to identify the major components and accessory proteins that constitute the native VSOAC multimeric protein complex. Since the activation of cardiac Cl- channels can produce significant effects on action potential duration and automaticity, and are key regulators of cell volume homeostasis, these channels have important clinical significance for several myocardial diseases, including cardiac arrhythmias, myocardial ischemia, congestive heart failure and hypertrophy. This project has significant potential of elucidating the normal physiological and possible pathophysiological role of volume-sensitive Cl- channels in the heart and cardiovascular system.