An evolutionary-conserved redox regulatory mechanism in human Ser/Thr protein kinases
An evolutionary-conserved redox regulatory mechanism in human Ser/Thr protein kinases
复制标题
DOI：
10.1101/571844
复制
发表时间：
2019-03
期刊：
影响因子：
--
通讯作者：
D. Byrne;Safal Shrestha;N. Kannan;P. Eyers
中科院分区：
文献类型：
--
作者：
D. Byrne;Safal Shrestha;N. Kannan;P. Eyers
活性氧（reactive oxygen species，ROS）是氧代谢的产物，也被认为是细胞信号传导的内源性生理介质。真核蛋白激酶（ePK）调节通过柔性激活片段中的可逆磷酸化事件发生。在这项研究中，我们证明，从有丝分裂的丝氨酸/苏氨酸激酶极光A的催化磷酸转移酶的输出也控制半胱氨酸（Cys）氧化。可逆的调节发生直接修改的保守残基（Cys 290），这是相邻的Thr 288，磷酸化的激活位点。引人注目的是，在其他ePKs中Cys 290-等价物的氧化还原调节被预测为未被充分认识的调节机制，因为约100种人类Ser/Thr激酶在保守激活环中的该位置处具有Cys。使用实时酶分析，我们证实，存在的等效半胱氨酸残基是预后的氧化还原敏感性之间的一组人类CAMK，AGC和AGC样激酶，包括AKT，AMPK，CAMK 1，MAPKAP-K2/3和SIK 1 -3。我们的研究结果表明，占主导地位的半胱氨酸为基础的氧化还原开关的激活段代表了一个进化保守的模式，为人类激酶组的一个重要子集的调节。这一发现对于理解ROS的生理和病理信号反应具有重要意义，并强调了ePKs中多价激活片段调控的重要性。Ser/Thr激酶的催化活性通过保守的基于Cys的氧化还原机制来调节。
Reactive oxygen species (ROS) are products of oxygen metabolism, but are also recognized as endogenous physiological mediators of cellular signaling. Eukaryotic protein kinase (ePK) regulation occurs through reversible phosphorylation events in the flexible activation segment. In this study, we demonstrate that the catalytic phosphotransferase output from the mitotic Ser/Thr kinase Aurora A is also controlled by cysteine (Cys) oxidation. Reversible regulation occurs by direct modification of a conserved residue (Cys 290), which lies adjacent to Thr 288, the activating site of phosphorylation. Strikingly, redox modulation of the Cys 290-equivalent in other ePKs is predicted to be an underappreciated regulatory mechanism, since ~100 human Ser/Thr kinases possess a Cys at this position in the conserved activation loop. Using real-time enzyme assays, we confirm that the presence of the equivalent Cys residue is prognostic for redox-sensitivity amongst a cohort of human CAMK, AGC and AGC-like kinases, including AKT, AMPK, CAMK1, MAPKAP-K2/3 and SIK1-3. Our findings demonstrate that dominant Cys-based redox-switching in the activation segment represents an evolutionary-conserved mode of regulation for a significant subset of the human kinome. This finding has important implications for understanding physiological and pathological signaling responses to ROS, and emphasises the importance of multivalent activation segment regulation in ePKs. ONE-SENTENCE SUMMARY The catalytic activity of Ser/Thr kinases is regulated through a conserved Cys-based redox mechanism.