Role of phosphorylation in cardiac muscle myosin

磷酸化在心肌肌球蛋白中的作用

• 批准号: 8746718
• 负责人: James Sellers
• 金额: $ 23.63万
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8746718
• 关键词: Address; Biological Assay; Cardiac; Cardiac Myosins; Goals; Heart; Heart Diseases; Human; In Vitro; Isometric Exercise; Left; Light; Measurement; Mechanics; Muscle; Myocardial; Myocardium; Myosin Light Chain Kinase; Myosin Regulatory Light Chains; Output; Performance; Phosphorylation; Physiological; Play; Property; Protein Kinase; Rattus; Rattus norvegicus; Recombinants; Role; Sampling; Series; Speed; Up-Regulation; Ventricular; Work; cell motility; non-muscle myosin; optical traps

Understanding how cardiac myosin regulatory light chain (RLC) phosphorylation alters cardiac muscle mechanics is important because it is often altered in cardiac disease. The effect this protein phosphorylation has on muscle mechanics during a physiological range of shortening velocities, during which the heart generates power and performs work, has not been addressed. We have expressed and phosphorylated recombinant Rattus norvegicus left ventricular RLC. In vitro we have phosphorylated these recombinant species with cardiac myosin light chain kinase and zipper-interacting protein kinase. We compare rat permeabilized cardiac trabeculae, which have undergone exchange with differently phosphorylated RLC species. We were able to enrich trabecular RLC phosphorylation by 40% compared with controls and, in a separate series, lower RLC phosphorylation to 60% of control values. Compared with the trabeculae with a low level of RLC phosphorylation, RLC phosphorylation enrichment increased isometric force by more than 3-fold and peak power output by more than 7-fold and approximately doubled both maximum shortening speed and the shortening velocity that generated peak power. We augmented these measurements by observing increased RLC phosphorylation of human and rat HF samples from endocardial left ventricular homogenate. These results demonstrate the importance of increased RLC phosphorylation in the up-regulation of myocardial performance and suggest that reduced RLC phosphorylation is a key aspect of impaired contractile function in the diseased myocardium. We are currently using optical trapping and in vitro motility assays to explore the effect of phosphorylation on purified cardiac muscle myosin.

了解心肌肌球蛋白调节轻链（RLC）磷酸化如何改变心肌力学是很重要的，因为它经常在心脏疾病中改变。这种蛋白质磷酸化在缩短速度的生理范围内对肌肉力学的影响尚未得到解决，在此期间，心脏产生动力并做功。我们表达并磷酸化重组褐家鼠左心室RLC。在体外，我们磷酸化这些重组物种与心肌肌球蛋白轻链激酶和拉链相互作用蛋白激酶。我们比较大鼠透化心脏小梁，这已经经历了不同的磷酸化RLC物种交换。与对照组相比，我们能够富集小梁RLC磷酸化40%，并且在单独的系列中，将RLC磷酸化降低至对照值的60%。与低水平RLC磷酸化的小梁相比，RLC磷酸化富集使等长收缩力增加了3倍以上，峰值功率输出增加了7倍以上，最大缩短速度和产生峰值功率的缩短速度增加了约一倍。我们通过观察来自内皮细胞左心室匀浆的人和大鼠HF样品的RLC磷酸化增加来增强这些测量。这些结果表明，增加RLC磷酸化的心肌性能的上调的重要性，并表明，减少RLC磷酸化是在患病心肌收缩功能受损的一个关键方面。 我们目前正在使用光学捕获和体外运动试验，以探讨磷酸化对纯化的心肌肌球蛋白的影响。