Structure/Function Studies on the Ca2+: Cation Antiporter family of transporters

Ca2 的结构/功能研究：阳离子逆向转运蛋白家族转运蛋白

基本信息

批准号：
7658647
负责人：
Jeffrey S Abramson
金额：
$ 23.1万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-01 至 2011-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7658647
关键词：
Animals Arrhythmia Bacteria Bioinformatics Biological Assay Brain Calcium Calcium Signaling Calcium-Binding Domain Cardiac Cardiac Myocytes Cations Cell physiology Cells Complex Crystallization Data Data Set Disease Drug Design Family Family member Goals Health Heart Heart failure Homeostasis Homologous Gene Ions Kidney Knowledge Lead Length Maintenance Membrane Proteins Methodology Monitor Muscle Cells Muscle relaxation phase Myocardium Nucleic Acid Regulatory Sequences Pharmacologic Substance Phase Physiology Plants Protein Family Protein Isoforms Proteins Regulation Reproducibility Resolution Robotics Role Sampling Signal Pathway Signal Transduction Structure System Technology Testing Tissues Transmembrane Domain Transport Process Yeasts antiporter cell type design insight member novel prevent public health relevance relating to nervous system three dimensional structure vector

项目摘要

DESCRIPTION (provided by applicant): The calcium-cation antiporter (CaCA) family of proteins is important for the control of intracellular calcium signaling. Maintenance of Ca2+ homeostasis depends on the functioning of specific transport systems that regulate Ca2+. One member of this family, the cardiac sarcolemmal Na+-Ca2+ exchanger (NCX1), is the primary mechanism for Ca2+ efflux from myocytes. As such, NCX1 is intimately involved in cardiac muscle relaxation and in preventing cellular Ca2+ overload. NCX1 has also been implicated in the genesis of some cardiac arrhythmias and has increased function in heart failure. This study will generate mechanistic details of a medically relevant protein from the CaCA family of exchangers, NCX1. In addition, we will develop new methodology for the expression, purification, crystallization and phase determination of the CaCA family. This technology will be directly transferable to other families of membrane proteins to yield high-resolution structures. In particular, we aim to achieve a 3D structure of the intracellular regulatory loop of NCX1, the primary regulator of Ca2+ extrusion from mammalian cardiac myocytes, as well as the structure of a complete exchanger from a prokaryotic member of the CaCA family. All hypotheses from the structural analysis will be tested by functional assays. The knowledge from these structure/function studies may lead to rational drug design and to the resolution of remaining unknown aspects of the function of this critical regulator of cardiac and neural physiology. PUBLIC HEALTH RELEVANCE: The CaCA family of proteins are ubiquitous and vital components of Ca2+ signaling pathways utilized for maintaining Ca2+ homeostasis in a variety of cell types. The mammalian Na+-Ca2+ exchanger (NCX), the best- characterized member of the family, is intimately involved in cardiac muscle relaxation and in preventing cellular Ca2+ overload. Determining the structure and regulatory function of CaCA family members in general, and NCX1 in particular, will allow for rational design of novel pharmaceuticals that may be used in regulating cardiac contractility in health and disease related to dysfunctional Ca2+ transport.

描述（由申请人提供）：钙阳离子逆向转运蛋白（CaCA）家族对于细胞内钙信号传导的控制很重要。 Ca2+ 稳态的维持取决于调节 Ca2+ 的特定运输系统的功能。该家族的一个成员是心脏肌膜 Na+-Ca2+ 交换器 (NCX1)，它是 Ca2+ 从肌细胞流出的主要机制。因此，NCX1 与心肌松弛和防止细胞 Ca2+ 超载密切相关。 NCX1 还与一些心律失常的发生有关，并且在心力衰竭中具有增强的功能。这项研究将产生来自 CaCA 交换蛋白家族的医学相关蛋白 NCX1 的机制细节。此外，我们还将开发CaCA家族表达、纯化、结晶和相测定的新方法。该技术将直接转移到其他膜蛋白家族，以产生高分辨率的结构。特别是，我们的目标是获得 NCX1 的细胞内调节环的 3D 结构，NCX1 是哺乳动物心肌细胞排出 Ca2+ 的主要调节因子，以及来自 CaCA 家族原核成员的完整交换器的结构。结构分析中的所有假设都将通过功能分析进行测试。这些结构/功能研究的知识可能会导致合理的药物设计，并解决心脏和神经生理学关键调节器功能中剩余的未知方面。公共健康相关性：CaCA 蛋白家族是 Ca2+ 信号通路中普遍存在的重要组成部分，用于维持多种细胞类型中的 Ca2+ 稳态。哺乳动物Na+-Ca2+交换器(NCX)是该家族中特征最鲜明的成员，与心肌松弛和防止细胞Ca2+超载密切相关。确定 CaCA 家族成员的结构和调节功能，特别是 NCX1，将有助于合理设计新型药物，这些药物可用于调节与 Ca2+ 转运功能障碍相关的健康和疾病中的心肌收缩力。