Skip to main content

Research Repository

Advanced Search

Spectroscopic and computational insight into the activation of O2 by the mononuclear Cu center in polysaccharide monooxygenases

Kjaergaard, Christian H.; Qayyum, Munzarin F.; Wong, Shaun D.; Xu, Feng; Hemsworth, Glyn R.; Walton, Daniel J.; Young, Nigel A.; Davies, Gideon J.; Walton, Paul H.; Johansen, Katja Salomon; Hodgson, Keith O.; Hedman, Britt; Solomon, Edward I.

Authors

Christian H. Kjaergaard

Munzarin F. Qayyum

Shaun D. Wong

Feng Xu

Glyn R. Hemsworth

Daniel J. Walton

Profile Image

Dr Nigel Young N.A.Young@hull.ac.uk
Senior Lecturer/ Director of Studies/ Deputy Head of Chemistry and Biochemistry/ Industrial Placements Coordinator

Gideon J. Davies

Paul H. Walton

Katja Salomon Johansen

Keith O. Hodgson

Britt Hedman

Edward I. Solomon



Abstract

Strategies for O₂ activation by copper enzymes were recently expanded to include mononuclear Cu sites, with the discovery of the copper-dependent polysaccharide monooxygenases, also classified as auxiliary-activity enzymes 9–11 (AA9-11). These enzymes are finding considerable use in industrial biofuel production. Crystal structures of polysaccharide monooxygenases have emerged, but experimental studies are yet to determine the solution structure of the Cu site and how this relates to reactivity. From X-ray absorption near edge structure and extended X-ray absorption fine structure spectroscopies, we observed a change from four-coordinate Cu(II) to three-coordinate Cu(I) of the active site in solution, where three protein-derived nitrogen ligands coordinate the Cu in both redox states, and a labile hydroxide ligand is lost upon reduction. The spectroscopic data allowed for density functional theory calculations of an enzyme active site model, where the optimized Cu(I) and (II) structures were consistent with the experimental data. The O₂ reactivity of the Cu(I) site was probed by EPR and stopped-flow absorption spectroscopies, and a rapid one-electron reduction of O₂ and regeneration of the resting Cu(II) enzyme were observed. This reactivity was evaluated computationally, and by calibration to Cu-superoxide model complexes, formation of an end-on Cu-AA9-superoxide species was found to be thermodynamically favored. We discuss how this thermodynamically difficult one-electron reduction of O₂ is enabled by the unique protein structure where two nitrogen ligands from His1 dictate formation of a T-shaped Cu(I) site, which provides an open coordination position for strong O₂ binding with very little reorganization energy.

Citation

Kjaergaard, C. H., Qayyum, M. F., Wong, S. D., Xu, F., Hemsworth, G. R., Walton, D. J., …Solomon, E. I. (2014). Spectroscopic and computational insight into the activation of O2 by the mononuclear Cu center in polysaccharide monooxygenases. Proceedings of the National Academy of Sciences of the United States of America, 111(24), 8797-8802. https://doi.org/10.1073/pnas.1408115111

Journal Article Type Article
Acceptance Date May 6, 2014
Online Publication Date Jun 2, 2014
Publication Date Jun 17, 2014
Deposit Date Feb 18, 2016
Publicly Available Date Feb 18, 2016
Journal Proceedings of the national academy of sciences
Print ISSN 0027-8424
Electronic ISSN 1091-6490
Publisher National Academy of Sciences
Peer Reviewed Peer Reviewed
Volume 111
Issue 24
Pages 8797-8802
DOI https://doi.org/10.1073/pnas.1408115111
Keywords X-ray absorption spectroscopy, DFT, Dioxygen activation, Biofuels
Public URL https://hull-repository.worktribe.com/output/471711
Publisher URL http://www.pnas.org/content/111/24/8797
Copyright Statement © 2016 National Academy of Sciences
Additional Information This is an author's accepted manuscript of an article published in Proceedings of the national academy of sciences, 2014, v.111 issue 24.

Files

Article.pdf (506 Kb)
PDF

Copyright Statement
© 2016 National Academy of Sciences





You might also like



Downloadable Citations