Lu Chang, Liu Xin, Zhang Chen-Song, Gong Haipeng, Wu Jia-Wei, Wang Zhi-Xin
Key Laboratory of Ministry of Education for Protein Science, School of Life Sciences, Tsinghua University , Beijing 100084, PR China.
State Key Laboratory of Stress Cell Biology, School of Life Sciences, Xiamen University , Xiamen, Fujian 361005, PR China.
Biochemistry. 2017 Nov 21;56(46):6165-6175. doi: 10.1021/acs.biochem.7b00827. Epub 2017 Nov 8.
The mitogen-activated protein kinases (MAPKs) are key components of cellular signal transduction pathways, which are down-regulated by the MAPK phosphatases (MKPs). Catalytic activity of the MKPs is controlled both by their ability to recognize selective MAPKs and by allosteric activation upon binding to MAPK substrates. Here, we use a combination of experimental and computational techniques to elucidate the molecular mechanism for the ERK2-induced MKP3 activation. Mutational and kinetic study shows that the FNFM motif in the MKP3 catalytic domain is essential for MKP3-mediated ERK2 inactivation and is responsible for ERK2-mediated MKP3 activation. The long-term molecular dynamics (MD) simulations further reveal a complete dynamic process in which the catalytic domain of MKP3 gradually changes to a conformation that resembles an active MKP catalytic domain over the time scale of the simulation, providing a direct time-dependent observation of allosteric signal transmission in ERK2-induced MKP3 activation.
丝裂原活化蛋白激酶(MAPKs)是细胞信号转导途径的关键组成部分,其受到MAPK磷酸酶(MKPs)的负调控。MKPs的催化活性既受其识别选择性MAPKs能力的控制,也受与MAPK底物结合后的变构激活作用的控制。在此,我们结合实验和计算技术来阐明ERK2诱导的MKP3激活的分子机制。突变和动力学研究表明,MKP3催化结构域中的FNFM基序对于MKP3介导的ERK2失活至关重要,并且是ERK2介导的MKP3激活的原因。长期分子动力学(MD)模拟进一步揭示了一个完整的动态过程,在该过程中,MKP3的催化结构域在模拟的时间尺度上逐渐转变为类似于活性MKP催化结构域的构象,提供了ERK2诱导的MKP3激活中变构信号传递的直接时间依赖性观察结果。