18O 动态同位素效应揭示了酸盐脱酶催化酸转移的关联过渡状态
Graeme W Howe1,2, Wilfred A van der Donk1,2,3
1Department of Chemistry , University of Illinois at Urbana-Champaign , 600 South Mathews Avenue , Urbana , Illinois 61801 , United States.
Journal of the American Chemical Society
|December 12, 2018
概括
酸脱酶 (PTDH) 使用一种关联机制来转移醇,与其他酸盐加工酶不同. 这涉及显著的核友性参与和紧密的过渡状态,由于贫穷的化物离开组.
科学领域:
- 生物化学 生物化学
- 酶动力学 酶动力学
- 生物有机化学 生物有机化学
背景情况:
- 酸脱酶 (PTDH) 催化了一种独特的酸转移反应,其中涉及化物离开组.
- 尽管有大量的研究努力,PTDH的催化机制 (关联性与分离性) 仍然不太清楚.
研究的目的:
- 通过描述其过渡状态 (TS) 来阐明热稳定PTDH (17X-PTDH) 的催化机制.
- 为了确定PTDH催化是否通过关联或离散途径进行.
主要方法:
- 确定初级2H和二级18O动态同位素效应 (KIEs).
- 在酶活性部位内的各种过渡状态结构的计算建模.
- 预测2H和18OKIEs以验证计算过渡状态.
主要成果:
- 大型,正常的18O KIEs强烈表明了关联性催化机制.
- 计算分析支持一个紧密的过渡状态与大量的核友参与.
- 观察到的机制与典型的酸盐单一和二反应的分离过渡状态形成鲜明对比.
结论:
- 17X-PTDH通过一种关联机制催化醇转移.
- 酶的过渡状态的特点是有限的离开组移位和显著的核友性攻击.
- 这种独特的机制归因于化物的离开组能力较差,需要形成强大的PO键.
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