Crystal Structures of Cyclohexanone Monooxygenase Reveal Complex Domain Movements and a Sliding Cofactor
Abstract
Cyclohexanone monooxygenase (CHMO) is a flavoprotein that carries out the archetypical Baeyer-Villiger oxidation of a variety of cyclic ketones into lactones. Using NADPH and O{sub 2} as cosubstrates, the enzyme inserts one atom of oxygen into the substrate in a complex catalytic mechanism that involves the formation of a flavin-peroxide and Criegee intermediate. We present here the atomic structures of CHMO from an environmental Rhodococcus strain bound with FAD and NADP+ in two distinct states, to resolutions of 2.3 and 2.2 {angstrom}. The two conformations reveal domain shifts around multiple linkers and loop movements, involving conserved arginine 329 and tryptophan 492, which effect a translation of the nicotinamide resulting in a sliding cofactor. Consequently, the cofactor is ideally situated and subsequently repositioned during the catalytic cycle to first reduce the flavin and later stabilize formation of the Criegee intermediate. Concurrent movements of a loop adjacent to the active site demonstrate how this protein can effect large changes in the size and shape of the substrate binding pocket to accommodate a diverse range of substrates. Finally, the previously identified BVMO signature sequence is highlighted for its role in coordinating domain movements. Taken together, these structures provide mechanistic insights into CHMO-catalyzed Baeyer-Villigermore »
- Authors:
- Publication Date:
- Research Org.:
- Brookhaven National Lab. (BNL), Upton, NY (United States). National Synchrotron Light Source
- Sponsoring Org.:
- Doe - Office Of Science
- OSTI Identifier:
- 980113
- Report Number(s):
- BNL-93031-2010-JA
Journal ID: ISSN 0002-7863; JACSAT; TRN: US201015%%1498
- DOE Contract Number:
- DE-AC02-98CH10886
- Resource Type:
- Journal Article
- Journal Name:
- Journal of the American Chemical Society
- Additional Journal Information:
- Journal Volume: 131; Journal Issue: 25; Journal ID: ISSN 0002-7863
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 36 MATERIALS SCIENCE; ARGININE; ATOMS; CRYSTAL STRUCTURE; CYCLOHEXANONE; ENZYMES; ISOALLOXAZINES; KETONES; LACTONES; NADP; NICOTINAMIDE; OXIDATION; OXYGEN; PROTEINS; RANGE; RHODOCOCCUS; SHAPE; SIZE; STRAINS; SUBSTRATES; TRYPTOPHAN; national synchrotron light source
Citation Formats
Mirza, I, Yachnin, B, Wang, S, Grosse, S, Bergeron, H, Imura, A, Iwaki, H, Hasegawa, Y, Lau, P, and Berghuis, A. Crystal Structures of Cyclohexanone Monooxygenase Reveal Complex Domain Movements and a Sliding Cofactor. United States: N. p., 2009.
Web. doi:10.1021/ja9010578.
Mirza, I, Yachnin, B, Wang, S, Grosse, S, Bergeron, H, Imura, A, Iwaki, H, Hasegawa, Y, Lau, P, & Berghuis, A. Crystal Structures of Cyclohexanone Monooxygenase Reveal Complex Domain Movements and a Sliding Cofactor. United States. https://doi.org/10.1021/ja9010578
Mirza, I, Yachnin, B, Wang, S, Grosse, S, Bergeron, H, Imura, A, Iwaki, H, Hasegawa, Y, Lau, P, and Berghuis, A. 2009.
"Crystal Structures of Cyclohexanone Monooxygenase Reveal Complex Domain Movements and a Sliding Cofactor". United States. https://doi.org/10.1021/ja9010578.
@article{osti_980113,
title = {Crystal Structures of Cyclohexanone Monooxygenase Reveal Complex Domain Movements and a Sliding Cofactor},
author = {Mirza, I and Yachnin, B and Wang, S and Grosse, S and Bergeron, H and Imura, A and Iwaki, H and Hasegawa, Y and Lau, P and Berghuis, A},
abstractNote = {Cyclohexanone monooxygenase (CHMO) is a flavoprotein that carries out the archetypical Baeyer-Villiger oxidation of a variety of cyclic ketones into lactones. Using NADPH and O{sub 2} as cosubstrates, the enzyme inserts one atom of oxygen into the substrate in a complex catalytic mechanism that involves the formation of a flavin-peroxide and Criegee intermediate. We present here the atomic structures of CHMO from an environmental Rhodococcus strain bound with FAD and NADP+ in two distinct states, to resolutions of 2.3 and 2.2 {angstrom}. The two conformations reveal domain shifts around multiple linkers and loop movements, involving conserved arginine 329 and tryptophan 492, which effect a translation of the nicotinamide resulting in a sliding cofactor. Consequently, the cofactor is ideally situated and subsequently repositioned during the catalytic cycle to first reduce the flavin and later stabilize formation of the Criegee intermediate. Concurrent movements of a loop adjacent to the active site demonstrate how this protein can effect large changes in the size and shape of the substrate binding pocket to accommodate a diverse range of substrates. Finally, the previously identified BVMO signature sequence is highlighted for its role in coordinating domain movements. Taken together, these structures provide mechanistic insights into CHMO-catalyzed Baeyer-Villiger oxidation.},
doi = {10.1021/ja9010578},
url = {https://www.osti.gov/biblio/980113},
journal = {Journal of the American Chemical Society},
issn = {0002-7863},
number = 25,
volume = 131,
place = {United States},
year = {Thu Jan 01 00:00:00 EST 2009},
month = {Thu Jan 01 00:00:00 EST 2009}
}