cGMP 依赖性蛋白激酶的晶体结构揭示了链间通讯的新位点。
Crystal structure of cGMP-dependent protein kinase reveals novel site of interchain communication.
机构信息
Department of Pharmacology, College of Medicine, University of Vermont, Burlington, VT 05405, USA.
出版信息
Structure. 2011 Sep 7;19(9):1317-27. doi: 10.1016/j.str.2011.06.012.
Abstract
The cGMP-dependent protein kinase (PKG) serves as an integral component of second messenger signaling in a number of biological contexts including cell differentiation, memory, and vasodilation. PKG is homodimeric and large conformational changes accompany cGMP binding. However, the structure of PKG and the molecular mechanisms associated with protomer communication following cGMP-induced activation remain unknown. Here, we report the 2.5 Å crystal structure of a regulatory domain construct (aa 78-355) containing both cGMP binding sites of PKG Iα. A distinct and segregated architecture with an extended central helix separates the two cGMP binding domains. Additionally, a previously uncharacterized helical domain (switch helix) promotes the formation of a hydrophobic interface between protomers. Mutational disruption of this interaction in full-length PKG implicates the switch helix as a critical site of dimer communication in PKG biology. These results offer new structural insight into the mechanism of allosteric PKG activation.
摘要
cGMP 依赖性蛋白激酶（PKG）作为第二信使信号转导的一个组成部分，在许多生物学背景下发挥作用，包括细胞分化、记忆和血管舒张。PKG 是同二聚体的，cGMP 结合会伴随构象的巨大变化。然而，PKG 的结构以及 cGMP 诱导激活后与亚基间通讯相关的分子机制仍然未知。在这里，我们报告了一个包含 PKG Iα 的两个 cGMP 结合位点的调节域结构（aa 78-355）的 2.5Å 晶体结构。一个独特且分隔的架构，带有一个延伸的中央螺旋，将两个 cGMP 结合域分开。此外，一个以前未被描述的螺旋域（开关螺旋）促进了亚基之间形成一个疏水界面。全长 PKG 中这种相互作用的突变破坏表明，开关螺旋是 PKG 生物学中亚基通讯的关键部位。这些结果为别构 PKG 激活的机制提供了新的结构见解。
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