Structures of carboxylic acid reductase reveal domain dynamics underlying catalysis

Deepanker Gahloth, Mark S Dunstan, Daniela Quaglia, Evaldas Klumbys, Michael P Lockhart-Cairns, Andrew Hill, Sasha R Derrington, Nigel S Scrutton, Nicholas J Turner, David Leys

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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.

Original languageEnglish
Pages (from-to)975-981
Number of pages7
JournalNature chemical biology
Volume13
Early online date17 Jul 2017
DOIs
Publication statusPublished - 2017

Keywords

  • Journal Article

Research Beacons, Institutes and Platforms

  • Manchester Institute of Biotechnology

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