Zhang Jiajing, King Christopher A, Dalby Kevin, Ren Pengyu
Department of Biomedical Engineering, The University of Texas at Austin, Austin, TX 78712, USA.
PMC Biophys. 2010 Jan 21;3(1):2. doi: 10.1186/1757-5036-3-2.
TRPM7/ChaK1 is a recently discovered atypical protein kinase that has been suggested to selectively phosphorylate the substrate residues located in alpha-helices. However, the actual structure of kinase-substrate complex has not been determined experimentally and the recognition mechanism remains unknown. In this work we explored possible kinase-substrate binding modes and the likelihood of an alpha-helix docking interaction, within a kinase active site, using molecular modeling. Specifically kinase ChaK1 and its two peptide substrates were examined; one was an 11-residue segment from the N-terminal domain of annexin-1, a putative endogenous substrate for ChaK1, and the other was an engineered 16-mer peptide substrate determined via peptide library screening. Simulated annealing (SA), replica-exchange molecular dynamics (REMD) and steered molecular dynamics (SMD) simulations were performed on the two peptide substrates and the ChaK1-substrate complex in solution. The simulations indicate that the two substrate peptides are unlikely to bind and react with the ChaK1 kinase in a stable alpha-helical conformation overall. The key structural elements, sequence motifs, and amino acid residues in the ChaK1 and their possible functions involved in the substrate recognition are discussed.PACS Codes: 87.15.A-
TRPM7/ChaK1是一种最近发现的非典型蛋白激酶,有人认为它能选择性地磷酸化位于α螺旋中的底物残基。然而,激酶-底物复合物的实际结构尚未通过实验确定,其识别机制仍然未知。在这项工作中,我们使用分子建模探索了激酶活性位点内可能的激酶-底物结合模式以及α螺旋对接相互作用的可能性。具体研究了激酶ChaK1及其两种肽底物;一种是来自膜联蛋白-1 N端结构域的11个残基片段,它是ChaK1的一种假定内源性底物,另一种是通过肽库筛选确定的工程化16聚体肽底物。对溶液中的两种肽底物以及ChaK1-底物复合物进行了模拟退火(SA)、副本交换分子动力学(REMD)和引导分子动力学(SMD)模拟。模拟结果表明,这两种底物肽总体上不太可能以稳定的α螺旋构象与ChaK1激酶结合并反应。讨论了ChaK1中的关键结构元件、序列基序和氨基酸残基及其在底物识别中可能涉及的功能。
87.15.A-