The effect on structural and solvent water molecules of substrate binding to ferric horseradish peroxidase.

The effect on structural and solvent water molecules of substrate binding to ferric horseradish peroxidase.
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对与辣根铁过氧化物酶结合的底物的结构和溶剂水分子的影响。

DOI:
10.1039/c4fd00161c
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发表时间:
2015
影响因子:
3.4
通讯作者:
Simpson N
Simpson N
中科院分区:
化学2区
文献类型:
--
作者:
Simpson N

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超快,多维红外光谱,在2D-IR和泵探测测量的形式,已被用来研究底物结合的辣根过氧化物酶(HRP)酶的结构动力学的影响。使用一氧化氮绑定到铁血红素的HRP作为一个敏感的探针的本地动力学,我们报告的测量的频率波动(光谱扩散)和振动寿命的NO伸缩模式与苯甲氧肟酸(BHA)位于基板结合的位置在周边的血红素口袋,在D2 O和H2O溶剂。结果表明,与BHA绑定的酶,局部结构动力学不敏感的H/D交换。这些结果与无底物酶研究中发现的结果形成鲜明对比,后者表明血红素配体的局部化学和动态环境受水分子的影响。鉴于底物结合引起的溶剂可及性的巨大变化,我们讨论了在不同阶段沿着酶促机制的反应坐标的HRP血红素口袋中的溶剂的不同作用的潜力。
Ultrafast, multi-dimensional infrared spectroscopy, in the form of 2D-IR and pump–probe measurements, has been employed to investigate the effect of substrate binding on the structural dynamics of the horseradish peroxidase (HRP) enzyme. Using nitric oxide bound to the ferric haem of HRP as a sensitive probe of local dynamics, we report measurements of the frequency fluctuations (spectral diffusion) and vibrational lifetime of the NO stretching mode with benzohydroxamic acid (BHA) located in the substrate-binding position at the periphery of the haem pocket, in both D2O and H2O solvents. The results reveal that, with BHA bound to the enzyme, the local structural dynamics are insensitive to H/D exchange. These results are in stark contrast to those found in studies of the substrate-free enzyme, which demonstrated that the local chemical and dynamic environment of the haem ligand is influenced by water molecules. In light of the large changes in solvent accessibility caused by substrate binding, we discuss the potential for varying roles for the solvent in the haem pocket of HRP at different stages along the reaction coordinate of the enzymatic mechanism.
辣根过氧化物酶与几种过氧化物反应的基本步骤:化合物0和化合物I形成的动力学和热力学
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