Regulatory Interactions of Cardiac Ion Pumps

描述（由申请人提供）：申请人提出了一个有两个目标的策略来探索离子驱动atp酶调节的分子机制。目的1关注心脏钙泵（SERCA）及其受磷蛋白（PLB）的调控。本研究将利用荧光共振能量转移（FRET）和化学交联来验证PLB与SERCA上多个位点结合的假设，以及这些离散的结合位点介导不同的功能作用。目的2描述了一种新的高通量分析方法，用于研究磷蛋白（PLM）与自身及其调控靶点钠/钾泵（NKA）的结合相互作用。该实验的目标是开发一种新型肽NKA超抑制剂，作为心脏糖苷（一类用于治疗心力衰竭的肌力药物）的更安全替代品。本应用程序的两个目的中描述的实验将为离子动机atp酶调节的基本机制提供新的见解，并可能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.