SIGNIFICANCE OF TROPONIN C MUTATIONS IN HEART DISEASE

肌钙蛋白 C 突变在心脏病中的意义

• 批准号: 7955893
• 负责人: CHEE CHEW LIM
• 金额: $ 0.14万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7955893
• 关键词: Amino Acids; Biological Models; Biology; Calcium; Calcium Binding; Calcium-Binding Domain; Cardiac; Cardiomyopathies; Computer Retrieval of Information on Scientific Projects Database; Development; Digestion; Funding; Gel; Genetic; Grant; Heart; Heart Diseases; Human; Institution; Link; Mass Spectrum Analysis; Medicine; Microfilaments; Missense Mutation; Mutation; Myocardial; Myocardial Contraction; Myocardial dysfunction; Paper; Patients; Peptides; Proteins; Relative (related person); Research; Research Personnel; Resources; Signal Transduction; Source; Structure-Activity Relationship; Troponin; Troponin C; United States National Institutes of Health; base; design; human disease; mutant; novel; tandem mass spectrometry; transmission process

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. Emerging evidence has linked mutations in myofiliment proteins to the development of genetic cardiomyopathies and myocardial dysfunction. Troponin C is the protein responsible for the transmission of the calcium-binding signal and triggering the contractile cycle. Dr. Liao and her associates identified two novel missense mutations in human cardiac troponin C at amino acid residues 59 (E59D) and 75 (D75Y) from a patient with idiopathic dialated cardiomyopathy, the first identified mutation of troponin C from any human disease. These missense mutations are located within the calcium-binding domain that regulates myocardial contraction, and result in decreased myofilament calcium responsiveness. These results showed mutations in troponin C contribute to the decreased contractile function in the diseased human heart. To determine the structure-function relationship, they designed a number of troponin C mutants based on replacing specific amino acid residues located within regulatory calcium-binding domains. We used in-gel proteolytic digestion, followed by MALDI-TOF and tandem mass spectrometry to structurally verify specific mutations. Additionally, semiquantitation of synthetic mutant peptides enabled estimation of relative expression levels. This mutational model system is helping to define how mutations in cardiac troponin C alter calcium responsiveness in cardiac myofilaments and, consequently, determine myocardial contractility. The paper describing these results was featured on the cover of Biophys. J. in May 2008.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 新的证据表明肌丝蛋白的突变与遗传性心肌病和心肌功能障碍的发生有关。肌钙蛋白 C 是负责传递钙结合信号并触发收缩周期的蛋白质。 Liao 博士和她的同事在一名特发性扩张性心肌病患者的人类心肌肌钙蛋白 C 氨基酸残基 59 (E59D) 和 75 (D75Y) 处发现了两个新的错义突变，这是首次在任何人类疾病中发现肌钙蛋白 C 突变。这些错义突变位于调节心肌收缩的钙结合域内，并导致肌丝钙反应性降低。这些结果表明肌钙蛋白 C 的突变导致患病人类心脏的收缩功能下降。为了确定结构-功能关系，他们基于替换位于调节性钙结合域内的特定氨基酸残基，设计了许多肌钙蛋白 C 突变体。我们使用凝胶内蛋白水解消化，然后使用 MALDI-TOF 和串联质谱法从结构上验证特定突变。此外，合成突变肽的半定量可以估计相对表达水平。该突变模型系统有助于确定心肌肌钙蛋白 C 的突变如何改变心肌丝中的钙反应性，从而确定心肌收缩力。描述这些结果的论文登上了《Biophys》的封面。 J.，2008 年 5 月。