PROTEIN EXCHANGE TO STUDY MUSCLE FUNCTION AND DISEASE

通过蛋白质交换研究肌肉功能和疾病

• 批准号: 2909812
• 负责人: JOSEPH M CHALOVICH
• 金额: $ 12.83万
• 依托单位: EAST CAROLINA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至 2002-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2909812
• 关键词: X ray crystallography; actins; enzyme activity; gene mutation; intermolecular interaction; laboratory rabbit; method development; muscle function; muscle metabolism; myosins; protein metabolism; protein structure function; striated muscles; troponin

DESCRIPTION (Adapted from applicant's abstract): Regulation of muscle contraction is difficult to study because it involves changes in several proteins that are in a highly organized system. Biochemical studies provide detailed information of the components but at the loss of the organization. Fiber studies preserve the structure but give little information about the function of the components. By exchanging native regulatory proteins in a single fiber with proteins containing a probe, it is possible to study the function of single components within a fiber. Fluorescent probes on troponin and tropomyosin have been shown, in solution studies, to be sensitive to calcium binding or to the fraction of actin in the active configuration. When placed into single fibers, the fluorescent probes can be used to monitor the state of the regulatory apparatus during various mechanical perturbations. This allows one to follow individual regulatory proteins in the organized structure of the fiber and to answer many questions related to regulation. The method of exchange that is used is an improvement over older methods and leaves the mechanical properties of the fibers unchanged. The plan is to study the exchange process so that it may be optimized and extended it to other proteins in striated muscle. A number of fluorescent probes will be placed on different regions of the troponin complex and the fluorescence changes that occur in fibers will be correlated with those occurring in solution under controlled conditions. In addition to the introduction of fluorescent probes, the exchange process will be used to introduce other well characterized modified or mutant troponins into fibers to study the function of particular regions of troponin. The intent is to begin by examining the effect of the mutations of the regulatory apparatus found in patients with hypertrophic cardiomyopathy. This work is part of a collaborative effort with Dr. Bernhard Brenner (mechanical studies), Dr. Leepo C. Yu (structural studies) and Dr. Yi-der Chen (mathematical modeling).

描述（改编自申请人摘要）：肌肉调节 收缩很难研究，因为它涉及到几个方面的变化。 蛋白质是一个高度有序的系统。 生化研究提供了 详细信息的组成部分，但在损失的组织。 纤维研究保留了结构，但提供的信息很少。 组件的功能。 通过交换天然调节蛋白， 单纤维与含有探针的蛋白质，有可能研究 纤维内单个组分的功能。 荧光探针 在溶液研究中，肌钙蛋白和原肌球蛋白被证明是 对钙结合或对活性细胞中肌动蛋白的分数敏感， 配置. 当放入单根纤维中时，荧光探针可以 用于在各种过程中监视调节装置的状态 机械扰动 这使得一个人能够遵循个人的监管规定， 蛋白质在纤维的组织结构，并回答许多 与监管有关的问题。 使用的交换方法是 改进了旧的方法，并留下了机械性能的 纤维不变。 计划是研究交换过程，以便 并将其扩展到横纹肌中的其他蛋白质。 一些 将荧光探针置于肌钙蛋白的不同区域， 复合物和纤维中发生的荧光变化将与 而那些在受控条件下在溶液中发生。 此外 为了引入荧光探针，将使用交换过程 为了将其他充分表征的修饰或突变的肌钙蛋白引入到 研究肌钙蛋白特定区域的功能。 意图 是开始通过检查基因突变的影响， 在肥厚型心肌病患者身上发现的器官 这项工作是 与Bernhard Brenner博士（机械）合作的一部分 研究），Dr. Leepo C. Yu（结构研究）和Yi-der Chen博士 （数学模型）。