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Publication Abstract from PubMed

Prenylated flavin mononucleotide (prFMN) is a modified flavin cofactor required by the UbiD family of (de)carboxylase enzymes. While the reduced prFMNH(2) form is produced by the flavin prenyltransferase UbiX, the corresponding two-electron oxidized prFMN(iminium) form is required to support UbiD catalysis. Thus, oxidative maturation of prFMNH(2) is required, which can be catalyzed by UbiD. However, heterologous (over)expression of UbiDs frequently leads to the accumulation of the stable but non-active one-electron oxidized purple prFMN(radical) species. A dedicated prFMN maturase enzyme (PhdC) from Mycolicibacterium fortuitum was recently identified as capable of catalyzing the oxidative maturation of prFMN(radical) to prFMN(iminium), thereby enabling an effective supply of active cofactor to the associated phenazine-1-carboxylate (de)carboxylase PhdA. We report the crystal structure of PhdC in complex with flavin, revealing it is a distant member of the class I HpaC-like family of short-chain dimeric flavin reductases and demonstrate catalytic conversion of the prFMN(radical) species to prFMN(iminium) in the presence of oxygen or ferricyanide. Co-expression of PhdC or a distant homologue from Priestia megaterium (YclD) with the canonical UbiD from Escherichia coli leads to activation of the latter, similar in effect to co-expression with the prFMNH(2)-binding chaperone LpdD. Conserved Glu residues in the PhdC active site suggest catalysis occurs through C1' proton-abstraction coupled oxidation. This study thus provides both structural and mechanistic insight into the function of PhdC, adding to the expanding repertoire of prFMN-binding proteins associated with the widespread UbiDX system.

Structure and Mechanism of PhdC, a Prenylated-Flavin Maturase.,Whittall DR, Box HG, Payne KAP, Marshall SA, Leys D Proteins. 2025 Dec 9. doi: 10.1002/prot.70096. PMID:41363046^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.