In vivo evidence for glycyl radical insertion into a catalytically inactive variant of pyruvate formate-lyase.

ABSTRACT: The dimeric glycyl radical enzyme pyruvate formate-lyase (PflB; formate acetyltransferase 1) cleaves pyruvate with hypothetical half-site reactivity to formate and acetyl-CoA. The radical introduced onto residue G734 of PflB is transiently transferred to C419 of an adjacent cysteine pair (C418/C419) during catalysis, but it is unclear whether glycyl radical formation is dependent on C419 in vivo. We show here that a deficiency in formate production of an Escherichia coli strain synthesizing a PflB_G734A variant, but lacking the autonomous glycyl radical cofactor, GrcA, could be restored by reintroducing plasmid-encoded native PflB, but not by a PflB_C418A/C419A variant, indicating that PflB_C418A/C419A cannot replace GrcA. Oxygen-dependent polypeptide cleavage

SUBMITTER: Kammel M 

PROVIDER: S-EPMC12338860 | biostudies-literature | 2025 May

REPOSITORIES: biostudies-literature