PMID:11444864

Citation	Wagner, AF, Schultz, S, Bomke, J, Pils, T, Lehmann, WD and Knappe, J (2001) YfiD of Escherichia coli and Y06I of bacteriophage T4 as autonomous glycyl radical cofactors reconstituting the catalytic center of oxygen-fragmented pyruvate formate-lyase. Biochem. Biophys. Res. Commun. 285:456-62
Abstract	Reaction of oxygen with the glycyl radical in pyruvate formate-lyase (PFL) leads to cleavage of the polypeptide backbone between N-Calpha of Gly734. A recombinant protein comprising the core of PFL (Ser1-Ser733) is shown here to associate with the YfiD protein (14 kDa) of Escherichia coli and likewise with the homologous T4 encoded Y06I protein, yielding upon reaction with PFL activase a heterooligomeric PFL enzyme that has full catalytic activity (35 U/nmol). Treatment of the activated complexes with oxygen led to cleavage of the 14 kDa proteins into 11 and 3 kDa polypeptides as expected for the localization of the putative glycyl radical at Gly102 (YfiD) or Gly95 (Y06I). For the isolated fragments from Y06I, mass spectrometric analysis (nanoESI-MS) determined a C-terminal serine carboxamide in the 11 kDa fragment, and a N-terminal oxalyl modification in the 3 kDa fragment. We speculate that YfiD in E. coli and other facultative anaerobic bacteria has evolved as a "spare part" for PFL's glycyl radical domain, utilized for rapid recovery of PFL activity (and thus ATP generation) in cells that have experienced oxidative stress.
Links	PubMed Online version:10.1006/bbrc.2001.5186
Keywords	Acetyltransferases/chemistry; Acetyltransferases/metabolism; Adenosine Triphosphate/metabolism; Amino Acid Sequence; Bacterial Proteins/chemistry; Bacterial Proteins/metabolism; Bacteriophage T4/metabolism; Escherichia coli/metabolism; Free Radicals; Glycine; Kinetics; Models, Chemical; Models, Molecular; Molecular Sequence Data; Molecular Weight; Oxidative Stress; Peptide Fragments/chemistry; Peptide Fragments/metabolism; Protein Structure, Secondary; Recombinant Proteins/chemistry; Recombinant Proteins/metabolism; Sequence Alignment; Sequence Homology, Amino Acid; Viral Proteins/chemistry; Viral Proteins/metabolism
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