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Title: Long-Range Electrostatics-Induced Two-Proton Transfer Captured by Neutron Crystallography in an Enzyme Catalytic Site

Abstract

Neutron crystallography was used to directly locate two protons before and after a pH-induced two-proton transfer between catalytic aspartic acid residues and the hydroxy group of the bound clinical drug darunavir, located in the catalytic site of enzyme HIV-1 protease. The two-proton transfer is triggered by electrostatic effects arising from protonation state changes of surface residues far from the active site. The mechanism and pH effect are supported by quantum mechanics/molecular mechanics (QM/MM) calculations. The low-pH proton configuration in the catalytic site is deemed critical for the catalytic action of this enzyme and may apply more generally to other aspartic proteases. Neutrons therefore represent a superb probe to obtain structural details for proton transfer reactions in biological systems at a truly atomic level.

Authors:
 [1];  [1];  [2];  [3];  [1];  [1];  [1];  [4];  [5];  [6];  [1];  [3];  [1]
+ Show Author Affiliations
  1. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  2. BARC, Mumbai (India)
  3. Georgia State Univ., Atlanta, GA (United States)
  4. Rutherford Appleton Lab., Didcot (United Kingdom)
  5. Institut Laue Langevin, Grenoble Cedex (France)
  6. National Institutes of Health, Bethesda, MD (United States)
Publication Date:
Research Org.:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1255684
Grant/Contract Number:  
AC05-00OR22725
Resource Type:
Accepted Manuscript
Journal Name:
Angewandte Chemie (International Edition)
Additional Journal Information:
Journal Name: Angewandte Chemie (International Edition); Journal Volume: 55; Journal Issue: 16; Journal ID: ISSN 1433-7851
Publisher:
Wiley
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Gerlits, Oksana, Wymore, Troy, Das, Amit, Shen, Chen -Hsiang, Parks, Jerry M., Smith, Jeremy C., Weiss, Kevin L., Keen, David A., Blakeley, Matthew P., Louis, John M., Langan, Paul, Weber, Irene T., and Kovalevsky, Andrey. Long-Range Electrostatics-Induced Two-Proton Transfer Captured by Neutron Crystallography in an Enzyme Catalytic Site. United States: N. p., 2016. Web. doi:10.1002/anie.201509989.
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Gerlits, Oksana, Wymore, Troy, Das, Amit, Shen, Chen -Hsiang, Parks, Jerry M., Smith, Jeremy C., Weiss, Kevin L., Keen, David A., Blakeley, Matthew P., Louis, John M., Langan, Paul, Weber, Irene T., & Kovalevsky, Andrey. Long-Range Electrostatics-Induced Two-Proton Transfer Captured by Neutron Crystallography in an Enzyme Catalytic Site. United States. https://doi.org/10.1002/anie.201509989
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Gerlits, Oksana, Wymore, Troy, Das, Amit, Shen, Chen -Hsiang, Parks, Jerry M., Smith, Jeremy C., Weiss, Kevin L., Keen, David A., Blakeley, Matthew P., Louis, John M., Langan, Paul, Weber, Irene T., and Kovalevsky, Andrey. Wed . "Long-Range Electrostatics-Induced Two-Proton Transfer Captured by Neutron Crystallography in an Enzyme Catalytic Site". United States. https://doi.org/10.1002/anie.201509989. https://www.osti.gov/servlets/purl/1255684.
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@article{osti_1255684,
title = {Long-Range Electrostatics-Induced Two-Proton Transfer Captured by Neutron Crystallography in an Enzyme Catalytic Site},
author = {Gerlits, Oksana and Wymore, Troy and Das, Amit and Shen, Chen -Hsiang and Parks, Jerry M. and Smith, Jeremy C. and Weiss, Kevin L. and Keen, David A. and Blakeley, Matthew P. and Louis, John M. and Langan, Paul and Weber, Irene T. and Kovalevsky, Andrey},
abstractNote = {Neutron crystallography was used to directly locate two protons before and after a pH-induced two-proton transfer between catalytic aspartic acid residues and the hydroxy group of the bound clinical drug darunavir, located in the catalytic site of enzyme HIV-1 protease. The two-proton transfer is triggered by electrostatic effects arising from protonation state changes of surface residues far from the active site. The mechanism and pH effect are supported by quantum mechanics/molecular mechanics (QM/MM) calculations. The low-pH proton configuration in the catalytic site is deemed critical for the catalytic action of this enzyme and may apply more generally to other aspartic proteases. Neutrons therefore represent a superb probe to obtain structural details for proton transfer reactions in biological systems at a truly atomic level.},
doi = {10.1002/anie.201509989},
journal = {Angewandte Chemie (International Edition)},
number = 16,
volume = 55,
place = {United States},
year = {Wed Mar 09 00:00:00 EST 2016},
month = {Wed Mar 09 00:00:00 EST 2016}
}
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https://doi.org/10.1002/anie.201509989
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Cited by: 38 works
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Figures / Tables:

Figure 1 Figure 1: Substrate-free HIV-1 protease showing catalytic Asp25 and Asp25’ residues and the lytic water molecule. Figure generated from PDB entry 2PC0.
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    • Kumar, Vijith; Pilati, Tullio; Quici, Silvio
    • Chemistry - A European Journal, Vol. 23, Issue 58
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    Drug Resistance Mutation L76V Alters Nonpolar Interactions at the Flap–Core Interface of HIV-1 Protease
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