A molecular study linking cTnT dynamics to genetic cardiomyopathy

将 cTnT 动力学与遗传性心肌病联系起来的分子研究

基本信息

批准号：
8061931
负责人：
STEVEN D SCHWARTZ
金额：
$ 40.11万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-12-15 至 2015-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8061931
关键词：
Actins Address Animals Basic Science Binding Binding Proteins Biological Biological Process Biophysics Calcium Binding Cardiac Cardiomyopathies Cereals Chemicals Complex Cyclic AMP-Dependent Protein Kinases Disease Distant Environment Exertion Familial Hypertrophic Cardiomyopathy Fiber Functional disorder Generations Genetic Health Heart Heart Diseases Hereditary Disease In Vitro Individual Induced Mutation Lead Link Location Measurement Mediating Methodology Microfilaments Microscopic Modeling Molecular Motor Movement Muscle Muscle Fibers Muscle function Mutate Mutation Myocardium Phosphorylation Phosphorylation Site Phosphotransferases Physiological Physiology Plant Roots Play Property Protein Biochemistry Proteins Research Site Skin Structure Thin Filament Tissues Tropomyosin Troponin Troponin T Vertebral column Vocabulary cell motility disease-causing mutation human disease inorganic phosphate movie mutant programs reconstitution research study response troponin-tropomyosin complex

项目摘要

DESCRIPTION (provided by applicant): Familial Hypertrophic Cardiomyopathy is a common and often devastating genetic cardiac disease. Specific mutations in cardiac proteins have been identified as the root cause of this disease, but they often exert their biological effect far from the site of mutation. Such effects, usually known collectively as allostery are part of the common vocabulary of protein biochemistry, and implementation of our research program will demonstrate how such effects are also part of a multi-protein controlling component of the cardiac motor - the thin filament. In particular we focus on Ca2+ binding to cTnT (long known to be a major component in the control of a beating heart,) and phosphorylation at a known important location in cTnI. The fact that the mutations we plan to study (all in cTnT) have in some cases been shown to effect significant changes on both these control mechanisms seems to demonstrate the principle of "action at a distance" but what is lacking is a translational understanding of how these changes cause disease from the molecular level to whole animal physiology. Allostery in a complex multi-component machine investigated in this fashion is thus both of great impact in basic science and of the highest significance in understanding the root cause of a devastating and relatively common human disease. To address these questions we have devised a research strategy of methodologies that range from computation on an all atom model of the troponin complex, tropomyosin, and an actin backbone, to biophysical measurements of the properties of wildtype and mutated reconstituted thin filaments, to fiber studies. The methodologies yield partially complementary yet overlapping information that provides a fully integrated analysis of this complex question. In order to better understand allostery in the function of these biological control agents in both health and disease we will study the following 2 specific aims: Specific Aim 1: To evaluate the molecular mechanism of the transduction of Ca2+ binding to the movement of tropomyosin and how this regulates the biophysics and physiology of the thin filament control of cardiac function in wildtype and known FHC-linked TNT1 mutations. Specific Aim 2: To evaluate the molecular mechanism of the phosphorylation of Ser 23/24 of cTnI in regulating myofilament activation in wildtype and known FHC-linked TNT1 mutations. PUBLIC HEALTH RELEVANCE: Familial Hypertrophic Cardiomyopathy is an often devastating and common cardiac genetic disease. Specific mutations in cardiac proteins have been identified as the root cause of this disease, but they often exert their biological effect far from the site of mutation. This application is aimed at elucidating how this "action at a distance" causes dysfunction and eventual human disease.

描述（由申请人提供）：家族性心肌病是一种常见且经常毁灭性的遗传心脏病。心脏蛋白中的特异性突变已被确定为该疾病的根本原因，但它们通常远离突变部位的生物学作用。这种效应通常被称为变构是蛋白质生物化学常见词汇的一部分，我们的研究计划的实施将证明这种影响也是心脏运动的多蛋白控制成分的一部分 - 薄丝。特别是，我们专注于Ca2+与CTNT的结合（长期以来已知是跳动心脏控制中的主要组成部分）和在CTNI中已知的重要位置的磷酸化。在某些情况下，我们计划研究的突变（全部在CTNT中）对这两种控制机制都会发生重大变化，这一事实似乎证明了“远距离行动”的原理，但是缺乏对这些变化如何从分子水平导致整个动物生理学导致疾病的转化理解。因此，以这种方式调查的复杂多组分机器中的变构对基础科学的影响很大，并且在理解毁灭性和相对常见的人类疾病的根本原因方面具有最高的意义。为了解决这些问题，我们制定了一种方法论的研究策略，该研究策略范围从肌钙蛋白复合物，肌动蛋白和肌动蛋白骨架的所有原子模型上的计算范围，到对野生型的性质的生物物理测量以及突变的重新建造的薄丝细丝，再到纤维研究。这些方法产生了部分互补但重叠的信息，该信息提供了对这个复杂问题的完全集成分析。为了更好地理解这些生物控制剂在健康和疾病中的功能上的变构，我们将研究以下两个特定目的：具体目的1：评估Ca2+结合型肌动蛋白的转导的分子机制，以及如何调节型薄丝纤维控制型在Wildtype和Fhcc-cncce and fhcc-link中的生物物理学和生理学的生理学。具体目的2：评估CTNI Ser 23/24磷酸化的分子机制，以调节野生型和已知的FHC链接TNT1突变的肌丝激活。公共卫生相关性：家族性心肌病经常是一种毁灭性和常见的心脏遗传疾病。心脏蛋白中的特异性突变已被确定为该疾病的根本原因，但它们通常远离突变部位的生物学作用。该应用旨在阐明这种“距离行动”如何引起功能障碍和最终的人类疾病。