ACTIVITY MODULATION OF PROTEIN KINASE AND PHOSPHATASE

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 磷酸化/去磷酸化是生物体中发生的最重要的化学反应，是调节细胞外效应物触发的许多不同生物事件的基础。我们的项目涉及确定两种蛋白激酶 AceK（也是一种蛋白磷酸酶）和 Etk，以及一种蛋白磷酸酶（钙调神经磷酸酶）的晶体结构。对于同时具有激酶和磷酸酶活性的AceK，将解析AceK的激酶和磷酸酶构象，以及AceK-ICDH复合物结构，以研究AceK活性开关和调节机制。将确定全长膜 Etk、其突变体及其与底物 Ugd 的复合物的结构。 对于钙调蛋白（真核生物中钙调蛋白激活的信号传导中央控制器），我们创建了一种融合构建体（称为 CBA），以促进钙调蛋白-钙调蛋白复合物的结构测定，该复合物代表钙调蛋白与完整底物复合物的结构。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Phosphorylation/dephosphorylation is the most crucial chemical reaction taking place in living organisms, which is the basis for the regulatory control of many diverse biological events triggered by extracellular effectors. Our project involves the determination of the crystal structures of two protein kinases, AceK (also a protein phosphatase) and Etk, and a protein phosphatase, calcineurin. For AceK, which possesses both kinase and phosphatase activities, the kinase and phosphatase conformations of AceK, as well as AceK-ICDH complex structures will be solved for investigating the AceK activity switch and regulatory mechanism. The structures of full length membrane Etk, its mutants, and its complex with substrate Ugd will be determined. For calcineurin, which is a calmodulin-activated central controller of signaling in eukaryotes, we have created a fusion construct (known as CBA) to facilitate the structure determination of calmodulin-calcineurin complex, which would represent a structure of calmodulin in complex with an intact substrate.