Deepankar Gahloth
Structures of carboxylic acid reductase reveal domain dynamics underlying catalysis
Gahloth, Deepankar; Dunstan, Mark S.; Quaglia, Daniela; Klumbys, Evaldas; Lockhart-Cairns, Michael P.; Hill, Andrew M.; Derrington, Sasha R.; Scrutton, Nigel S.; Turner, Nicholas J.; Leys, David
Authors
Mark S. Dunstan
Daniela Quaglia
Evaldas Klumbys
Michael P. Lockhart-Cairns
Andrew M. Hill
Sasha R. Derrington
Nigel S. Scrutton
Nicholas J. Turner
David Leys
Abstract
Carboxylic acid reductase (CAR) catalyzes the ATP- and NADPH-dependent reduction of carboxylic acids to the corresponding aldehydes. The enzyme is related to the nonribosomal peptide synthetases, consisting of an adenylation domain fused via a peptidyl carrier protein (PCP) to a reductase termination domain. Crystal structures of the CAR adenylation-PCP didomain demonstrate that large-scale domain motions occur between the adenylation and thiolation states. Crystal structures of the PCP-reductase didomain reveal that phosphopantetheine binding alters the orientation of a key Asp, resulting in a productive orientation of the bound nicotinamide. This ensures that further reduction of the aldehyde product does not occur. Combining crystallography with small-angle X-ray scattering (SAXS), we propose that molecular interactions between initiation and termination domains are limited to competing PCP docking sites. This theory is supported by the fact that (R)-pantetheine can support CAR activity for mixtures of the isolated domains. Our model suggests directions for further development of CAR as a biocatalyst.
Citation
Gahloth, D., Dunstan, M. S., Quaglia, D., Klumbys, E., Lockhart-Cairns, M. P., Hill, A. M., Derrington, S. R., Scrutton, N. S., Turner, N. J., & Leys, D. (2017). Structures of carboxylic acid reductase reveal domain dynamics underlying catalysis. Nature Chemical Biology, 13(9), 975-981. https://doi.org/10.1038/nchembio.2434
Journal Article Type | Article |
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Acceptance Date | Jun 8, 2017 |
Online Publication Date | Jul 17, 2017 |
Publication Date | Sep 1, 2017 |
Deposit Date | Jun 18, 2020 |
Publicly Available Date | Jul 7, 2020 |
Journal | Nature Chemical Biology |
Print ISSN | 1552-4450 |
Electronic ISSN | 1552-4469 |
Publisher | Nature Publishing Group |
Peer Reviewed | Peer Reviewed |
Volume | 13 |
Issue | 9 |
Pages | 975-981 |
DOI | https://doi.org/10.1038/nchembio.2434 |
Public URL | https://nottingham-repository.worktribe.com/output/4672184 |
Publisher URL | https://www.nature.com/articles/nchembio.2434 |
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