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(19)F核磁共振和密度泛函理论分析揭示了RhoA催化的GTP水解的结构和电子过渡态特征。

(19)F NMR and DFT Analysis Reveal Structural and Electronic Transition State Features for RhoA-Catalyzed GTP Hydrolysis.

作者信息

Jin Yi, Molt Robert W, Waltho Jonathan P, Richards Nigel G J, Blackburn G Michael

机构信息

Krebs Institute, Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield, S10 2TN, UK.

Department of Chemistry and Chemical Biology, Indiana University Purdue University Indianapolis, Indianapolis, IN, 46202, USA.

出版信息

Angew Chem Int Ed Engl. 2016 Mar 1;55(10):3318-22. doi: 10.1002/anie.201509477. Epub 2016 Jan 28.

DOI:10.1002/anie.201509477
PMID:26822702
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4770445/
Abstract

Molecular details for RhoA/GAP catalysis of the hydrolysis of GTP to GDP are poorly understood. We use (19)F NMR chemical shifts in the MgF3(-) transition state analogue (TSA) complex as a spectroscopic reporter to indicate electron distribution for the γ-PO3(-) oxygens in the corresponding TS, implying that oxygen coordinated to Mg has the greatest electron density. This was validated by QM calculations giving a picture of the electronic properties of the transition state (TS) for nucleophilic attack of water on the γ-PO3(-) group based on the structure of a RhoA/GAP-GDP-MgF3(-) TSA complex. The TS model displays a network of 20 hydrogen bonds, including the GAP Arg85' side chain, but neither phosphate torsional strain nor general base catalysis is evident. The nucleophilic water occupies a reactive location different from that in multiple ground state complexes, arising from reorientation of the Gln-63 carboxamide by Arg85' to preclude direct hydrogen bonding from water to the target γ-PO3(-) group.

摘要

RhoA/GAP催化GTP水解为GDP的分子细节尚不清楚。我们使用MgF3(-)过渡态类似物（TSA）复合物中的（19）F NMR化学位移作为光谱报告物，以指示相应过渡态中γ-PO3(-)氧原子的电子分布，这意味着与Mg配位的氧具有最大的电子密度。通过量子力学计算验证了这一点，该计算基于RhoA/GAP-GDP-MgF3(-) TSA复合物的结构，给出了水对γ-PO3(-)基团进行亲核攻击的过渡态（TS）的电子性质图。该TS模型显示了一个由20个氢键组成的网络，包括GAP Arg85'侧链，但磷酸盐扭转应变和一般碱催化均不明显。亲核水占据的反应位置与多个基态复合物中的不同，这是由于Gln-63羧酰胺通过Arg85'重新定向，从而阻止了水与目标γ-PO3(-)基团直接形成氢键。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76cc/4770445/2552676db852/ANIE-55-3318-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76cc/4770445/8e2f98700e13/ANIE-55-3318-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76cc/4770445/1cecba1520a1/ANIE-55-3318-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76cc/4770445/b5f2d2bf2155/ANIE-55-3318-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76cc/4770445/b690884bd343/ANIE-55-3318-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76cc/4770445/2552676db852/ANIE-55-3318-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76cc/4770445/8e2f98700e13/ANIE-55-3318-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76cc/4770445/1cecba1520a1/ANIE-55-3318-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76cc/4770445/b5f2d2bf2155/ANIE-55-3318-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76cc/4770445/b690884bd343/ANIE-55-3318-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76cc/4770445/2552676db852/ANIE-55-3318-g005.jpg

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