title: Alternative pyrimidine biosynthesis protein ApbE is a flavin transferase catalyzing covalent attachment of FMN to a threonine residue in bacterial flavoproteins.
abstract: Na(+)-translocating NADH:quinone oxidoreductase (Na(+)-NQR) contains two flavin residues as redox-active prosthetic groups attached by a phosphoester bond to threonine residues in subunits NqrB and NqrC. We demonstrate here that flavinylation of truncated Vibrio harveyi NqrC at Thr-229 in Escherichia coli cells requires the presence of a co-expressed Vibrio apbE gene. The apbE genes cluster with genes for Na(+)-NQR and other FMN-binding flavoproteins in bacterial genomes and encode proteins with previously unknown function. Experiments with isolated NqrC and ApbE proteins confirmed that ApbE is the only protein factor required for NqrC flavinylation and also indicated that the reaction is Mg(2+)-dependent and proceeds with FAD but not FMN. Inactivation of the apbE gene in Klebsiella pneumoniae, wherein the nqr operon and apbE are well separated in the chromosome, resulted in a complete loss of the quinone reductase activity of Na(+)-NQR, consistent with its dependence on covalently bound flavin. Our data thus identify ApbE as a novel modifying enzyme, flavin transferase.
authors.affiliation: Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119992, Russia.
authors.forename: Yulia V
authors.initials: YV
authors.lastname: Bertsova
authors.affiliation: Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119992, Russia.
authors.forename: Maria S
authors.initials: MS
authors.lastname: Fadeeva
authors.affiliation: Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119992, Russia.
authors.forename: Vitaly A
authors.initials: VA
authors.lastname: Kostyrko
authors.affiliation: Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119992, Russia.
authors.forename: Marina V
authors.initials: MV
authors.lastname: Serebryakova
authors.affiliation: Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119992, Russia.
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Baykov
authors.affiliation: Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119992, Russia. Electronic address: bogachev@genebee.msu.ru.
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Bogachev
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D002384:Catalysis; D004579:Electron Transport; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005486:Flavin Mononucleotide / Q000378:metabolism*; D005415:Flavins / Q000378:metabolism*; D005822:Genetic Vectors; D007711:Klebsiella pneumoniae / Q000201:enzymology* / Q000235:genetics; D008274:Magnesium / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000737:chemistry* / Q000378:metabolism; D009876:Operon; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D011743:Pyrimidines / Q000096:biosynthesis*; D017386:Sequence Homology, Amino Acid; D014733:Vibrio / Q000201:enzymology*
chemical_list: D005415:Flavins; D011743:Pyrimidines; D005486:Flavin Mononucleotide; D009713:Nucleotidyltransferases; D008274:Magnesium
keywords: Bacterial Metabolism; FMN; Flavoproteins; Na+-translocating NADH:Quinone Oxidoreductase; Post-translational Modification; Respiratory Chain
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