8AP0

ForT Mutant T138V

This is a non-PDB format compatible entry.

Summary for 8AP0

• Entry DOI: 10.2210/pdb8ap0/pdb
• Descriptor: Beta-ribofuranosylaminobenzene 5'-phosphate synthase, 1-O-pyrophosphono-5-O-phosphono-alpha-D-ribofuranose, 4-azanyl-1~{H}-pyrazole-3,5-dicarboxylic acid, ... (7 entities in total)
• Functional Keywords: formycin biosynthesis pathway protein c-nucleoside formation enzyme, structural protein
• Biological source: Streptomyces kaniharaensis
• Total number of polymer chains: 1
• Total formula weight: 37434.12

Authors

Li, W.,Naismith, J.H. (deposition date: 2022-08-09, release date: 2022-09-07, Last modification date: 2024-02-07)

Primary citation

Li, W.,Girt, G.C.,Radadiya, A.,Stewart, J.J.P.,Richards, N.G.J.,Naismith, J.H.
Experimental and computational snapshots of C-C bond formation in a C-nucleoside synthase.
Open Biology, 13:220287-220287, 2023

PubMed Abstract: The biosynthetic enzyme, ForT, catalyses the formation of a C-C bond between 4-amino-1-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.

PubMed: 36629016
DOI: 10.1098/rsob.220287

Experimental method

X-RAY DIFFRACTION (1.6 Å)