Glycine oxidase from Bacillus subtilis. Characterization of a new flavoprotein.
Glycine oxidase (GO) is a homotetrameric flavoenzyme that contains one molecule of non-covalently bound flavin adenine dinucleotide per 47 kDa protein monomer. GO is active on various amines (sarcosine, N-ethylglycine, glycine) and d-amino acids (d-alanine, d-proline). The products of GO reaction with various substrates have been determined, and it has been clearly shown that GO catalyzes the oxidative deamination of primary and secondary amines, a reaction similar to that of d-amino acid oxidase, although its sequence homology is higher with enzymes such as sarcosine oxidase and N-methyltryptophane oxidase. GO shows properties that are characteristic of the oxidase class of flavoproteins: it stabilizes the anionic flavin semiquinone and forms a reversible covalent flavin-sulfite complex. The approximately 300 mV separation between the two FAD redox potentials is in accordance with the high amount of the anionic semiquinone formed on photoreduction. GO can be distinguished from d-amino acid oxidase by its low catalytic efficiency and high apparent K(m) value for d-alanine. A number of active site ligands have been identified; the tightest binding is observed with glycolate, which acts as a competitive inhibitor with respect to sarcosine. The presence of a carboxylic group and an amino group on the substrate molecule is not mandatory for binding and catalysis.
Job
Viviana
V
Department of Structural and Functional Biology, University of Insubria, via J. H. Dunant 3, 21100 Varese, Italy.
Marcone
Giorgia Letizia
GL
Pilone
Mirella S
MS
Pollegioni
Loredano
L
D000445:Aldehyde Oxidoreductases / Q000378:metabolism; D000594:Amino Acid Oxidoreductases / Q000737:chemistry* / Q000502:physiology*; D001412:Bacillus subtilis / Q000201:enzymology*; D001665:Binding Sites; D002087:Butyrates / Q000737:chemistry; D002384:Catalysis; D018076:DNA, Complementary / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005420:Flavoproteins / Q000737:chemistry*; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007651:Keto Acids / Q000737:chemistry; D007700:Kinetics; D008024:Ligands; D008956:Models, Chemical; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity; D013447:Sulfites / Q000378:metabolism
D002087:Butyrates; D018076:DNA, Complementary; D005420:Flavoproteins; D007651:Keto Acids; D008024:Ligands; D013447:Sulfites; D000445:Aldehyde Oxidoreductases; C087006:formaldehyde dehydrogenase, glutathione-independent; D000594:Amino Acid Oxidoreductases; D005969:Glutamate Dehydrogenase; C115925:glycine oxidase; D010100:Oxygen