Department of Biological Sciences, University of Essex, Colchester, Essex, UK.
FEBS J. 2009 Dec;276(24):7305-18. doi: 10.1111/j.1742-4658.2009.07440.x.
The hypothesis is tested that Jun-Fos activator protein-1 coiled coil interactions are dominated during late folding events by the formation of intricate intermolecular electrostatic contacts. A previously derived cJun-FosW was used as a template as it is a highly stable relative of the wild-type cJun-cFos coiled coil protein (thermal melting temperature = 63 degrees C versus 16 degrees C), allowing kinetic folding data to be readily extracted. An electrostatic mutant, cJun(R)-FosW(E), was created to generate six Arg-Glu interactions at e-g'+1 positions between cJun(R) and FosW(E), and investigations into how their contribution to stability is manifested in the folding pathway were undertaken. The evidence now strongly indicates that the formation of interhelical electrostatic contacts exert their effect predominantly on the coiled coil unfolding/dissociation rate. This has major implications for future antagonist design whereby kinetic rules could be applied to increase the residency time of the antagonist-peptide complex, and therefore significantly increase the efficacy of the antagonist.
该假说经过验证，在晚期折叠事件中，Jun-Fos 激活蛋白-1 卷曲螺旋相互作用主要由复杂的分子间静电接触形成。先前推导的 cJun-FosW 被用作模板，因为它是野生型 cJun-cFos 卷曲螺旋蛋白的高度稳定相对物（热融解温度 = 63°C 与 16°C），允许轻易提取动力学折叠数据。创建了静电突变体 cJun(R)-FosW(E)，在 cJun(R)和 FosW(E)之间的 e-g'+1 位置产生六个 Arg-Glu 相互作用，并研究了它们对稳定性的贡献如何在折叠途径中表现出来。现在有强有力的证据表明，螺旋间静电接触的形成主要对卷曲螺旋展开/解离速率产生影响。这对未来的拮抗剂设计具有重大意义，通过动力学规则可以增加拮抗剂肽复合物的停留时间，从而显著提高拮抗剂的疗效。
