STRUCTURAL CHANGES IN SIGNALING IN CALCIUM REGULATIONS

描述(改编自申请人的摘要)： 拟议的研究是为了了解钙依赖的分子基础 对生化活动的调节。实验方法是利用 CaM与骨骼肌和平滑肌肌球蛋白轻链的相互作用 利用具有酶活性的截断突变体的蛋白激酶(MLCK)。这个 申请者将研究底物和三磷酸腺苷如何调节这些 相互作用，并寻找2Ca+.CaM.MLCK中间体的证据 失活的复合体。申请者将调查两国之间的互动 肌动蛋白肌动蛋白催化核心的纤维连接蛋白结构域N端 与催化核心的结合以及CaM结合到 MLCK。 CaM与磷酸化酶(PhK)的相互作用--利用CaM结合 来自催化亚单位PhK(302-306)的多肽。 CaM与血管平滑肌蛋白钙调蛋白(CAD)的相互作用 来自CAD序列的CaM结合肽。 TNC/TnI在骨骼肌和心肌形式中的相互作用，与 强调改进当前的骨骼相互作用模型和 理解功能差异的结构基础 心脏系统。申请者还将把这些研究扩展到三元制 TNC/TnI/TnT复合体使用心脏系统。 生化调节是健康功能的关键。监管细目 机制，如果没有纠正，通常会导致病理状况和 不受控制的增殖或细胞死亡。了解潜在的 因此，分子调控机制在生物多样性研究中具有重要意义 健康研究。

DESCRIPTION (Adapted from applicant's abstract): The objective of the proposed research is to understand the molecular basis for Ca2+-dependent regulation of biochemical activity. The experimental approach is to utilize CaM's interactions with both skeletal and smooth muscle myosin light chain kinase (MLCK) utilizing enzymatically active truncation mutants. The applicant will investigate how substrate and ATP modulate these interactions, and look for evidence of a 2Ca2+.CaM.MLCK intermediate inactivated complex. The applicant will investigate the interaction of the fibronectin domain N-terminal to the catalytic core in smooth muscle MLCK with the catalytic core and how it might be modulated by CaM binding to MLCK. CaM's interaction with phosphorylase kinase (PhK), utilizing CaM-binding peptides from the catalytic subunit PhK(302-306). CaM's interaction with the smooth muscle protein caldesmon (CaD) utilizing CaM-binding peptides from the CaD sequence. TnC/TnI interactions in both skeletal muscle and cardiac forms, with an emphasis on improving the current model for skeletal interactions and understanding the structural basis for the differences in function with the cardiac system. The applicant will also expand these studies to the ternary TnC/TnI/TnT complex using the cardiac system. Biochemical regulation is key to healthy function. Breakdown of regulatory mechanisms if uncorrected generally leads to pathological conditions and to uncontrolled proliferation or cell death. Understanding the underlying molecular mechanisms of regulation is thus of fundamental importance in health research.