Understanding the Specificity of a Docking Interaction between JNK1 and the Scaffolding Protein JIP1

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• 作者：Chunli Yan ; Tamer Kaoud ; Sunbae Lee ; Kevin N. Dalby ; Pengyu Ren
• 刊名：Journal of Physical Chemistry B
• 出版年：2011
• 出版时间：February 17, 2011
• 年：2011
• 卷：115
• 期：6
• 页码：1491-1502
• 全文大小：1306K
• 年卷期：v.115,no.6(February 17, 2011)
• ISSN：1520-5207

文摘

The up-regulation of JNK activity is associated with a number of disease states. The JNK鈭扟IP1 interaction represents an attractive target for the inhibition of JNK-mediated signaling. In this study, molecular dynamics simulations have been performed on the apo-JNK1 and the JNK1鈥?span class="smallcaps">l-pepJIP1 and JNK1鈥?span class="smallcaps">d-pepJIP1 complexes to investigate the interaction between the JIP1 peptides and JNK1. Dynamic domain studies based on essential dynamics (ED) analysis of apo-JNK1 and the JNK1鈥?span class="smallcaps">l-pepJIP1 complex have been performed to analyze and compare details of conformational changes, hinge axes, and hinge bending regions in both structures. The activation loop, the 伪C helix, and the G loop are found to be highly flexible and to exhibit significant changes in dynamics upon l-pepJIP1 binding. The conformation of the activation loop for the apo state is similar to that of inactive apo-ERK2, while the activation loop in JNK1鈥?span class="smallcaps">l-pepJIP1 complex resembles that of the inactive ERK2 bound with pepHePTP. ED analysis shows that, after the binding of l-pepJIP1, the N- and C-terminal domains of JNK1 display both a closure and a twisting motion centered around the activation loop, which functions as a hinge. In contrast, no domain motion is detected for the apo state for which an open conformation is favored. The present study suggests that l-pepJIP1 regulates the interdomain motions of JNK1 and potentially the active site via an allosteric mechanism. The binding free energies of l-pepJIP1 and d-pepJIP1 to JNK1 are estimated using the molecular mechanics Poisson鈭払oltzmann and generalized-Born surface area (MM-PB/GBSA) methods. The contribution of each residue at the interaction interface to the binding affinity of l-pepJIP1 with JNK1 has been analyzed by means of computational alanine-scanning mutagenesis and free energy decomposition. Several critical interactions for binding (e.g., Arg156/l-pepJIP1 and Glu329/JNK1) have been identified. The binding free energy calculation indicates that the electrostatic interaction contributes critically to specificity, rather than to binding affinity between the peptide and JNK1. Notably, the binding free energy calculations predict that d-pepJIP1 binding to JNK1 is significantly weaker than the L form, contradicting the previous suggestion that d-pepJIP1 acts as an inhibitor toward JNK1. We have performed experiments using purified JNK1 to confirm that, indeed, d-pepJIP1 does not inhibit the ability of JNK1 to phosphorylate c-Jun in vitro.