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Human Butyrylcholinesterase Mutants for (−)-Cocaine Hydrolysis: A Correlation Relationship between Catalytic Efficiency and Total Hydrogen Bonding Energy with an Oxyanion Hole
The Journal of Physical Chemistry B ( IF 2.8 ) Pub Date : 2023-12-08 , DOI: 10.1021/acs.jpcb.3c06392 Fang Zheng 1, 2 , Shurong Hou 1 , Liu Xue 1 , Wenchao Yang 1 , Chang-Guo Zhan 1, 2
The Journal of Physical Chemistry B ( IF 2.8 ) Pub Date : 2023-12-08 , DOI: 10.1021/acs.jpcb.3c06392 Fang Zheng 1, 2 , Shurong Hou 1 , Liu Xue 1 , Wenchao Yang 1 , Chang-Guo Zhan 1, 2
Affiliation
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A combined computational and experimental study has been carried out to explore and test a quantitative correlation relationship between the relative catalytic efficiency (RCE) of human butyrylcholinesrase (BChE) mutant-catalyzed hydrolysis of substrate (−)-cocaine and the total hydrogen bonding energy (tHBE) of the carbonyl oxygen of the substrate with the oxyanion hole of the enzyme in the modeled transition-state structure (TS1), demonstrating a satisfactory linear correlation relationship between ln(RCE) and tHBE. The satisfactory correlation relationship has led us to computationally predict and experimentally confirm new human BChE mutants that have a further improved catalytic activity against (−)-cocaine, including the most active one (the A199S/F227S/S287G/A328W/Y332G mutant) with a 2790-fold improved catalytic efficiency (kcat/KM = 2.5 × 109 min–1 M–1) compared to the wild-type human BChE. Compared to the reference mutant (the A199S/S287G/A328W/Y332G mutant) tested in the reported clinical development of an enzyme therapy for cocaine dependence treatment, this new mutant (with a newly predicted additional F227S mutation) has an improved catalytic efficiency against (−)-cocaine by ∼2.6-fold. The good agreement between the computational and experimental ln(RCE) values suggests that the obtained correlation relationship is robust for computational prediction. A similar correlation relationship could also be explored in studying BChE or other serine hydrolases/esterases with an oxyanion hole stabilizing the carbonyl oxygen in the rate-determining reaction step of the enzymatic hydrolysis of other substrates.
中文翻译：
用于 (−)-可卡因水解的人丁酰胆碱酯酶突变体：催化效率和含氧阴离子空穴的总氢键能之间的相关关系
我们进行了计算和实验相结合的研究，以探索和测试人丁酰胆碱酶（BChE）突变体催化底物（−）-可卡因水解的相对催化效率（RCE）与总氢键能之间的定量相关关系（模拟过渡态结构（TS1）中底物的羰基氧与酶的氧阴离子孔的tHBE），证明ln（RCE）和tHBE之间具有令人满意的线性相关关系。令人满意的相关关系使我们能够通过计算预测和实验证实新的人类 BChE 突变体，这些突变体对 (−)-可卡因具有进一步提高的催化活性，其中包括最活跃的突变体（A199S/F227S/S287G/A328W/Y332G 突变体）与野生型人类 BChE 相比，催化效率提高了 2790 倍（ k cat / K M = 2.5 × 10 9 min –1 M –1 ）。与已报道的用于可卡因依赖治疗的酶疗法临床开发中测试的参考突变体（A199S/S287G/A328W/Y332G 突变体）相比，这种新突变体（具有新预测的额外 F227S 突变）对（ −)-可卡因约2.6倍。计算和实验 ln(RCE) 值之间的良好一致性表明所获得的相关关系对于计算预测是稳健的。在研究 BChE 或其他具有氧阴离子孔的丝氨酸水解酶/酯酶时，也可以探索类似的相关关系，该氧阴离子孔在其他底物的酶促水解的限速反应步骤中稳定羰基氧。
更新日期：2023-12-08
中文翻译：
用于 (−)-可卡因水解的人丁酰胆碱酯酶突变体：催化效率和含氧阴离子空穴的总氢键能之间的相关关系
我们进行了计算和实验相结合的研究，以探索和测试人丁酰胆碱酶（BChE）突变体催化底物（−）-可卡因水解的相对催化效率（RCE）与总氢键能之间的定量相关关系（模拟过渡态结构（TS1）中底物的羰基氧与酶的氧阴离子孔的tHBE），证明ln（RCE）和tHBE之间具有令人满意的线性相关关系。令人满意的相关关系使我们能够通过计算预测和实验证实新的人类 BChE 突变体，这些突变体对 (−)-可卡因具有进一步提高的催化活性，其中包括最活跃的突变体（A199S/F227S/S287G/A328W/Y332G 突变体）与野生型人类 BChE 相比，催化效率提高了 2790 倍（ k cat / K M = 2.5 × 10 9 min –1 M –1 ）。与已报道的用于可卡因依赖治疗的酶疗法临床开发中测试的参考突变体（A199S/S287G/A328W/Y332G 突变体）相比，这种新突变体（具有新预测的额外 F227S 突变）对（ −)-可卡因约2.6倍。计算和实验 ln(RCE) 值之间的良好一致性表明所获得的相关关系对于计算预测是稳健的。在研究 BChE 或其他具有氧阴离子孔的丝氨酸水解酶/酯酶时，也可以探索类似的相关关系，该氧阴离子孔在其他底物的酶促水解的限速反应步骤中稳定羰基氧。
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