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Structures of carboxylic acid reductase reveal domain dynamics underlying catalysis

DOI: 10.1038/nchembio.2434 DOI Help

Authors: Deepanker Gahloth (University of Manchester) , Mark S. Dunstan (University of Manchester) , Daniela Quaglia (University of Manchester) , Evaldas Klumbys (University of Manchester) , Michael P. Lockhart-cairns (University of Manchester; Diamond Light Source) , Andrew M. Hill (University of Manchester) , Sasha R. Derrington (University of Manchester) , Nigel S. Scrutton (University of Manchester) , Nicholas J Turner (University of Manchester) , David Leys (University of Manchester)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Chemical Biology , VOL 1146

State: Published (Approved)
Published: July 2017
Diamond Proposal Number(s): 12788

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.

Journal Keywords: Enzyme mechanisms; Enzymes; X-ray crystallography

Subject Areas: Biology and Bio-materials, Chemistry

Instruments: B21-High Throughput SAXS , I04-Macromolecular Crystallography