title: X-ray structure of Escherichia coli pyridoxine 5'-phosphate oxidase complexed with FMN at 1.8 A resolution.
abstract: BACKGROUND
  Escherichia coli pyridoxine 5'-phosphate oxidase (PNPOx) catalyzes the terminal step in the biosynthesis of pyridoxal 5'-phosphate (PLP), a cofactor used by many enzymes involved in amino acid metabolism. The enzyme oxidizes either the 4'-hydroxyl group of pyridoxine 5'-phosphate (PNP) or the 4'-primary amine of pyridoxamine 5'-phosphate (PMP) to an aldehyde. PNPOx is a homodimeric enzyme with one flavin mononucleotide (FMN) molecule non-covalently bound to each subunit. A high degree of sequence homology among the 15 known members of the PNPOx family suggests that all members of this group have similar three-dimensional folds.
  
  
  RESULTS
  The crystal structure of PNPOx from E. coli has been determined to 1.8 A resolution. The monomeric subunit folds into an eight-stranded beta sheet surrounded by five alpha-helical structures. Two monomers related by a twofold axis interact extensively along one-half of each monomer to form the dimer. There are two clefts at the dimer interface that are symmetry-related and extend from the top to the bottom of the dimer. An FMN cofactor that makes interactions with both subunits is located in each of these two clefts.
  
  
  CONCLUSIONS
  The structure is quite similar to the recently deposited 2.7 A structure of Saccharomyces cerevisiae PNPOx and also, remarkably, shares a common structural fold with the FMN-binding protein from Desulfovibrio vulgaris and a domain of chymotrypsin. This high-resolution E. coli PNPOx structure permits predictions to be made about residues involved in substrate binding and catalysis. These predictions provide testable hypotheses, which can be answered by making site-directed mutants.
authors.affiliation: Institute for Structural Biology and Drug Discovery, Virginia Commonwealth University, Richmond, VA 23219, USA. msafo@hsc.vcu.edu
authors.forename: M K
authors.initials: MK
authors.lastname: Safo
authors.forename: I
authors.initials: I
authors.lastname: Mathews
authors.forename: F N
authors.initials: FN
authors.lastname: Musayev
authors.forename: M L
authors.initials: ML
authors.lastname: di Salvo
authors.forename: D J
authors.initials: DJ
authors.lastname: Thiel
authors.forename: D J
authors.initials: DJ
authors.lastname: Abraham
authors.forename: V
authors.initials: V
authors.lastname: Schirch
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D002918:Chymotrypsin / Q000737:chemistry; D018360:Crystallography, X-Ray; D016969:Desulfovibrio vulgaris / Q000201:enzymology; D019281:Dimerization; D004579:Electron Transport; D004926:Escherichia coli / Q000201:enzymology*; D005486:Flavin Mononucleotide / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010710:Phosphates / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011734:Pyridoxaminephosphate Oxidase / Q000737:chemistry* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
chemical_list: D001426:Bacterial Proteins; D010710:Phosphates; D011993:Recombinant Fusion Proteins; D005486:Flavin Mononucleotide; D011734:Pyridoxaminephosphate Oxidase; D002918:Chymotrypsin