Regulatory Interactions of Cardiac Ion Pumps

描述（由申请人提供）：申请人提出了一种双目标策略，以探索离子动力ATP酶调节的分子机制。目的1研究心脏钙泵（SERCA）及其受磷蛋白（PLB）的调节作用。拟议的研究将使用荧光共振能量转移（FRET）和化学交联来测试PLB与SERCA上的多个位点结合的假设，并且这些离散的结合位点介导不同的功能效应。目的2描述了一种新的高通量检测方法，用于研究磷脂酶与其自身及其调节靶点钠/钾泵（NKA）的结合相互作用。拟议的实验的目标是开发一种新的肽超级抑制剂NKA作为一种更安全的替代强心苷，一类用于治疗心力衰竭的正性肌力药物。本申请的两个目的中描述的实验将提供对离子运动ATP酶调节的基本机制的新见解，并且可以提高我们对与心力衰竭相关的离子转运障碍的理解。

DESCRIPTION (provided by applicant): The applicant proposes a two aim strategy to explore molecular mechanisms of the regulation of ion-motive ATPases. Aim 1 focuses on the cardiac calcium pump (SERCA) and its regulation by phospholamban (PLB). The proposed research will use fluorescence resonance energy transfer (FRET) and chemical crosslinking to test the hypothesis that PLB binds to multiple sites on SERCA, and that these discrete binding sites mediate different functional effects. Aim 2 describes a novel high throughput assay for investigation of the binding interactions of phospholemman (PLM) with itself and its regulatory target, the sodium/potassium pump (NKA). The goal of the proposed experiments are to develop a novel peptide superinhibitor of NKA as a safer alternative to cardiac glycosides, a class of inotropic drugs used in the treatment of heart failure. The experiments described in the two Aims of this application will provide new insight into fundamental mechanisms of regulation of ion-motive ATPases, and may improve our understanding of the ion transport disorders associated with heart failure.