骨架蛋白Cypher作为PKA的锚定蛋白参与扩张型心肌病发病机制的研究

扩张型心肌病（DCM）病因复杂，疗效差。骨架蛋白基因突变致功能失衡是引起DCM的重要机制，这为寻找合适的干预靶点提供了机会。目前明确骨架蛋白Cypher缺失可导致DCM。前期实验我们发现Cypher是一个蛋白激酶A（cyclic-AMP dependent protein kinase A，PKA）的锚定蛋白（A kinase anchoring protein, AKAP），模拟结构和体外实验证实其中与心肌病相关的T206I突变可增强Cypher与PKA的结合。AKAP可调控PKA和蛋白磷酸酶的活性，影响下游蛋白的磷酸化。本课题用Cypher基因敲除和T206I敲入小鼠，用分子生物学技术，磷酸化蛋白组学方法、酵母双杂交以及质谱分析，探索Cypher-PKA复合物如何调控下游蛋白的磷酸化及T206I突变后致病的分子机理，从而揭示Cypher与DCM发病的关系，为寻找DCM干预靶点提供依据。

Dilated cardiomyopathy(DCM) has complicated etiology and it's treatment is undesirable. Dysfunction on cytoskeleton proteins due to mutations is one of important mechanisms, which brings potential targets for therapy. It has been well demonstrated that Cypher is a DCM associated gene. Our previous study has investigated that Cypher is a novel A kinase anchoring protein (AKAP). Moreover, one of cardiomyopathy associated Cypher mutation T206I, enhanced the interaction between Cypher and PKA. AKAP is a scaffold protein for protein kinases and phosphatases to compartamentate their activities, therefore to fine regulate protein phosphorylation. Using Cypher deficient and T206I mutation knockin mouse models, together with phosphoproteomic and mass spectrometry, we will find new substrates for AKAP of Cypher and the molecular mechanisms how T206I causes dilated cardiomyopathy. Our results bring new molecular events to understand dilated cardiomyopathy, finally for best clilical treatments.