ELUCIDATION OF THE INTRACELLULAR CALCIUM SIGNAL TRANSDUCTION WITH THE MOLECULAR PHARMACOLOGICAL APPROARCH

细胞信号的研究在过去的10年里取得了突破性的进展，它依赖于生理、生化和酶的研究，并引入了分子生物学的方法，如克隆基因。我们从蛋白质化学、分子生物学和分子药理学的角度研究了细胞内钙信号转导系统的分子基础。我们研究了一种新的钙/钙调蛋白依赖的蛋白激酶V(CaM Kinase V)被磷酸化激活的机制。在此基础上，制备了抗CaM激酶V的多克隆抗体，并对其组织分布和免疫组织化学定位进行了研究。然后我们发现了CaM激酶V-激活剂，它是CaM激酶V催化活性所必需的。有趣的是，CaM激酶V激活剂确实使与其激活相关的钙/钙调蛋白依赖的蛋白激酶IV(CaM激酶IV)磷酸化。最后，我们推测可能存在新的CaM激酶级联，如MAP激酶级联。我们一直在研究利用合成的分子来激活或抑制靶分子的活性来分析生物功能。在这些研究的基础上，我们设计了作用于细胞内信号转导的分子探针，阐明了与细胞内信号转导密切相关的CaM激酶的生理功能。最近，我们从大鼠脑中克隆了两个新的钙/钙调蛋白依赖性蛋白激酶I(CaM Kinase I)亚型，而CaM Kinase V被认为是CaM Kinase I的一个亚型。

Research on cellular signals, which depend heavily on physiological, biochemical and enzymatic studies, has made a rapid progress in the last 10 years with the break through introduction of such molecular biotogical approach as cDNA cloning. We studied molecular basis of the intracellular calcium signal transduction system by protein chemical, molecular biological, and molecular pharmacological approach. We have examined the mechanisms by which a novel Ca2+/calmodulin-dependent protein kinase V(CaM kinase V) was activated by phosphorylation. Physicochemical or biochemical properties of CaM kinase V were also analyzed Furthermore, polyclonal antibody against CaM kinase V was prepared and tissue distribution and immunohistochemical localization of CaM kinase V were studied. We have then discovered the CaM kinase V-activator which was necessary for CaM kinase V catalytic activity. Interestingly, the CaM kinase V-activator did phosphorylate Ca2+/calmodulin-dependent protein kinase IV(CaM kinase IV)associated with its activation. Finally, we suggested that there were new CaM kinase cascades such as those of MAP kinase. We have been studying to analyze biological function using synthesized molecule to activate or inhibit the activities of the target molecule. Based on these studies in creating designed molecular probes acting in the intracellular signal transduction, we have clarified the physiological functions of the CaM kinase which are closely linked to the intracellular signal transduction. Recently, we have cloned two new Ca2+/calmodulin-dependent protein kinase I(CaM kinase I) isoforms from rat brain and CaM kinase V has been thought to be an isoform of CaM kinase I.