Experimental and computational snapshots of C-C bond formation in a C-nucleoside synthase

Li, Wenbo, Girt, Georgina C., Radadiya, Ashish ORCID: https://orcid.org/0000-0001-7348-1755, Stewart, James J. P., Richards, Nigel G. J. ORCID: https://orcid.org/0000-0002-0375-0881 and Naismith, James H. 2023. Experimental and computational snapshots of C-C bond formation in a C-nucleoside synthase. Open Biology 13 (1) , 220287. 10.1098/rsob.220287

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Abstract

The biosynthetic enzyme, ForT, catalyses the formation of a C-C bond between 4-amino-1H-pyrazoledicarboxylic acid and MgPRPP to produce a C-nucleoside precursor of formycin A. The transformation catalysed by ForT is of chemical interest because it is one of only a few examples in which C-C bond formation takes place via an electrophilic substitution of a small, aromatic heterocycle. In addition, ForT is capable of discriminating between the aminopyrazoledicarboxylic acid and an analogue in which the amine is replaced by a hydroxyl group; a remarkable feat given the steric and electronic similarities of the two molecules. Here we report biophysical measurements, structural biology and quantum chemical calculations that provide a detailed molecular picture of ForT-catalysed C-C bond formation and the conformational changes that are coupled to catalysis. Our findings set the scene for employing engineered ForT variants in the biocatalytic production of novel, anti-viral C-nucleoside and C-nucleotide analogues.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Advanced Research Computing @ Cardiff (ARCCA) Chemistry
Additional Information:	License information from Publisher: LICENSE 1: URL: http://creativecommons.org/licenses/by/4.0/, Type: open-access
Publisher:	The Royal Society
Funders:	BBSRC, EPSRC
Date of First Compliant Deposit:	12 January 2023
Date of Acceptance:	30 November 2022
Last Modified:	02 Aug 2024 08:31
URI:	https://orca.cardiff.ac.uk/id/eprint/155806

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