Conformational Dynamics of FERM-Mediated Autoinhibition in Pyk2 Tyrosine Kinase
Conformational Dynamics of FERM-Mediated Autoinhibition in Pyk2 Tyrosine Kinase
复制标题
Pyk2 酪氨酸激酶中 FERM 介导的自抑制的构象动力学
DOI：
10.1021/acs.biochem.9b00541
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发表时间：
2019
期刊：
影响因子：
2.9
通讯作者：
Underbakke, Eric S.
中科院分区：
文献类型：
--
作者：
Loving, Hanna S.;Underbakke, Eric S.
Pyk 2是一种非受体酪氨酸激酶，由粘着斑激酶（FAK）的基因复制和随后在脑和造血细胞中的功能特化进化而来。Pyk 2与FAK共享一个结构域组织，其N-末端调节FERM结构域邻接激酶结构域。FAK调节涉及整联蛋白介导的膜聚集，以减轻FERM和激酶结构域之间的自抑制相互作用。Pyk 2调控仍然是神秘的，涉及Ca 2+内流和蛋白质支架。虽然FAK FERM结构域在自抑制中的机制是公认的，但Pyk 2 FERM的调节作用是模糊的。我们使用氢/氘交换质谱和激酶活性分析来探讨FERM介导的Pyk 2自身抑制的机制。结果揭示了负责自抑制的FERM-激酶接口。Pyk 2自身抑制影响激活环构象。此外，自身抑制的FERM-激酶界面表现出与FAK和Pyk 2中保守的FERM基本补丁的变构连接。
Pyk2 is a non-receptor tyrosine kinase that evolved from gene duplication of focal adhesion kinase (FAK) and subsequent functional specialization in the brain and hemopoietic cells. Pyk2 shares a domain organization with FAK, with an N-terminal regulatory FERM domain adjoining the kinase domain. FAK regulation involves integrin-mediated membrane clustering to relieve autoinhibitory interactions between FERM and kinase domains. Pyk2 regulation remains cryptic, involving Ca2+influx and protein scaffolding. While the mechanism of the FAK FERM domain in autoinhibition is well-established, the regulatory role of the Pyk2 FERM is ambiguous. We probed the mechanisms of FERM-mediated autoinhibition of Pyk2 using hydrogen/deuterium exchange mass spectrometry and kinase activity profiling. The results reveal FERM–kinase interfaces that are responsible for autoinhibition. Pyk2 autoinhibition impacts the activation loop conformation. In addition, the autoinhibitory FERM–kinase interface exhibits allosteric linkage with the FERM basic patch conserved in both FAK and Pyk2.