Characterization of the Dielectric Constant in the Trichoderma reesei Cel7B Active Site

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• 作者：Xiangfei Song ; Yefei Wang ; Shujun Zhang ; Shihai Yan ; Tong Li ; Lishan Yao
• 刊名：Journal of Chemical Information and Modeling
• 出版年：2015
• 出版时间：July 27, 2015
• 年：2015
• 卷：55
• 期：7
• 页码：1369-1376
• 全文大小：369K
• ISSN：1549-960X

文摘

An attempt is made to evaluate the dielectric constant of the Trichoderma reesei Cel7B active site. Through kinetic measurements, the pK_a value of the catalytic acid E201 is determined. Mutations (away from E201) with net charge changes are introduced to perturb the E201 pK_a. It is shown that the mutation with a +1 charge change (including G225R, G230R, and A335R) decreases the pK_a of E201, whereas the mutation with a 鈭? charge change (including Q149E, A222D, G225D, and G230D) increases the pK_a. This effect is consistent with the electrostatic interaction between the changed charge and the E201 side chain. The fitting of the experimental data yields an apparent dielectric constant of 25鈥?0. Molecular dynamics simulations with explicit water molecules indicate that the high solvent accessibility of the active site contributes largely to the high dielectric constant. ONIOM calculations show that high dielectric constant benefits the catalysis through decreasing the energy of the transition state relative to that of the enzyme substrate complex.