在酶催化的“深度”氢隧穿反应中利用高压和二级同位素效应探究活性位点几何结构。
Probing active site geometry using high pressure and secondary isotope effects in an enzyme-catalysed 'deep' H-tunnelling reaction.
作者信息
Hay Sam, Pudney Christopher R, Sutcliffe Michael J, Scrutton Nigel S
机构信息
Manchester Interdisciplinary Biocentre and Faculty of Life Science, University of Manchester, 131 Princess Street, Manchester M1 7ND, UK.
出版信息
J Phys Org Chem. 2010 Jul 1;23(7):696-701. doi: 10.1002/poc.1653.
Abstract
We report the first study of the effects of hydrostatic pressure on α-2° KIEs for an enzyme-catalysed H-transfer reaction that occurs by 'deep' tunnelling. High pressure causes a significant decrease in the observed α-2° KIE on the pre-steady-state hydride transfer from NADH to FMN in the flavoprotein morphinone reductase. We have recently shown that high pressure causes a reduction in macroscopic reaction barrier width for this reaction. Using DFT vibrational analysis of a simple active site model, we posit that the decrease in α-2° KIE with pressure may arise due to a decrease in the vibrational coupling between the NADH primary (transferred) and secondary hydrogens in the 'tunnelling ready configuration', which more closely resembles the reactant state than the transition state.
摘要
我们报告了第一项关于流体静压力对通过“深度”隧穿发生的酶催化氢转移反应的α-2°动力学同位素效应（KIE）影响的研究。高压导致黄素蛋白吗啡酮还原酶中从NADH到FMN的稳态前氢化物转移过程中观察到的α-2°KIE显著降低。我们最近表明，高压会导致该反应的宏观反应势垒宽度减小。通过对一个简单活性位点模型进行密度泛函理论（DFT）振动分析，我们推测随着压力增加α-2°KIE降低可能是由于在“隧穿就绪构型”中NADH一级（转移）氢和二级氢之间的振动耦合减少，这种构型与反应物状态的相似性高于过渡态。
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