Regulation of Cardiac Calcium Channels by an Autoinhibitory Signaling Complex

描述（由申请人提供）：Cav1.2通道传导的L型Ca电流负责启动心肌收缩的Ca进入，因此这些通道是许多不同效应物（如神经递质、激素和药物及其受体）调节Ca信号传导和收缩力的最终共同途径。L型钙电流的改变与心血管疾病和治疗密切相关。L型钙电流的失调导致高血压，钙通道拮抗剂药物是治疗高血压的重要方式。缺血性心脏病常伴有心绞痛，也可采用钙拮抗剂药物治疗。心律失常可通过改变L型Ca电流的调节和不适当定时的Ca瞬变产生早期和延迟后去极化而产生，Ca拮抗剂药物在治疗房性心律失常中很重要。心力衰竭时L型Ca电流的β-肾上腺素能调节改变。令人惊讶的是，尽管它们在心血管生理学和病理生理学中的重要性，但心肌细胞中Cav1.2通道的调节尚未得到很好的理解。由于它们在收缩调节中的关键作用，许多细胞内调节剂和第二信使会聚在这些Ca通道上并调节它们的功能，包括Mg、cAMP、Ca和钙调蛋白。我们的工作表明，这些第二信使的作用位点是在大的细胞内C-末端结构域，这代表了约30%的质量的α 1亚基。此外，C-末端结构域经受蛋白水解加工，这调节其功能。因此，C-末端结构域整合了多种细胞调节信号，它们共同形成了控制Ca通道活性的整合的细胞内信号网络。本项目拟通过对Cav1.2通道C端结构域的自抑制作用的分子机制和生理意义的研究，明确Cav1.2通道C端蛋白水解加工的机制和生理意义，并确定通过β-肾上腺素能受体途径调节Cav1.2通道的分子机制，所述β-肾上腺素能受体途径通过PKA结合到通道的远端C-AKAP 15的末端结构域。我们的实验结果将是至关重要的了解钙和cAMP信号在心肌细胞的调节和心血管疾病的功能障碍。这些信息将为旨在预防和治疗心血管疾病的转化研究提供必要的基础科学背景。

DESCRIPTION (provided by applicant): L-type Ca currents conducted by Cav1.2 channels are responsible for Ca entry that initiates contraction in cardiac muscle, and these channels are therefore the final common pathway for regulation of Ca signaling and contractile force by many different effectors such as neurotransmitters, hormones and drugs, and their receptors. Alterations in L-type Ca currents are crucially involved in cardiovascular disease and therapy. Misregulation of L-type Ca currents contributes to hypertension, and Ca channel antagonist drugs are an important mode of therapy. Ischemic heart disease is often accompanied by angina pectoris, which is also treated with Ca antagonist drugs. Arrhythmias can be generated by altered regulation of L-type Ca currents and by inappropriately timed Ca transients generating early and delayed afterdepolarizations, and Ca antagonist drugs are important in treatment of atrial arrhythmias, (-adrenergic regulation of L-type Ca currents is altered in heart failure. Surprisingly, despite their importance in cardiovascular physiology and pathophysiology, regulation of Cav1.2 channels in cardiac myocytes is not well understood. Because of their key role in regulation of contraction, many intracellular regulators and second messengers converge on these Ca channels and regulate their function, including Mg, cAMP, Ca, and calmodulin. Our work has shown that the sites of action of these second messengers are in the large intracellular C-terminal domain, which represents approximately 30% of the mass of the al subunit. In addition, the C-terminal domain is subject to proteolytic processing, which modulates its function. Thus, the C-terminal domain integrates many kinds of cellular regulatory signals, which together form an integrated intracellular signaling network controlling Ca channel activity. In this project we propose to determine the molecular mechanism and physiological significance of Cav1.2 channel autoinhibition the C-terminal domain, define the mechanism and physiological significance of proteolytic processing of the C-terminal of Cav1.2 channels, and determine the molecular mechanism of regulation of the Cav1.2 channel by the (-adrenergic receptor pathway acting through PKA bound to the channel's distal C-terminal domain by AKAP15. The results of our experiments will be crucial for understanding regulation of Ca and cAMP signaling in the cardiac myocyte and its dysfunction in cardiovascular disease. This information will provide the essential basic science background for translational research aimed at preventing and treating cardiovascular disease.