Amino acid sequence of long chain alpha-hydroxy acid oxidase from rat kidney, a member of the family of FMN-dependent alpha-hydroxy acid-oxidizing enzymes.
The complete amino acid sequence of rat kidney long chain alpha-hydroxy acid oxidase has been determined by microsequencing, using a number of standard enzymatic and chemical cleavages. Peptides were purified by high pressure liquid chromatography or by gel electrophoresis followed by electrotransfer. The sequence comprises 352 residues and ends with a peroxisomal targeting sequence SRL. The present work definitely establishes that hydroxy acid oxidase is a member of the family of FMN-dependent alpha-hydroxy acid-oxidizing enzymes. The family includes lactate oxidase, short chain alpha-hydroxy acid oxidase (glycolate oxidase), flavocytochrome b2, and mandelate dehydrogenase. There are altogether 45 totally conserved positions among the six sequences known. The sequence similarities are analyzed in light of the known three-dimensional structure of flavocytochrome b2 and glycolate oxidase. It is concluded that long chain hydroxy acid oxidase should be folded as a beta 8 alpha 8 barrel and should dehydrogenate alpha-hydroxy acids according to the same chemical mechanism as other enzymes of the family, in spite of a Tyr----Phe substitution at the active site.
Diêp Lê
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Centre National de la Recherche Scientifique Unité de Recherche Associée 1461, Hôpital Necker, Paris, France.
Lederer
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D000429:Alcohol Oxidoreductases / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry; D001665:Binding Sites; D005949:Glucose Oxidase / Q000737:chemistry; D007668:Kidney / Q000201:enzymology*; D007770:L-Lactate Dehydrogenase / Q000737:chemistry; D042844:L-Lactate Dehydrogenase (Cytochrome); D006899:Mixed Function Oxygenases / Q000737:chemistry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D051381:Rats; D016415:Sequence Alignment
D001426:Bacterial Proteins; D010455:Peptides; D006899:Mixed Function Oxygenases; D000429:Alcohol Oxidoreductases; D007770:L-Lactate Dehydrogenase; D042844:L-Lactate Dehydrogenase (Cytochrome); C021380:L-2-hydroxyacid oxidase; D005949:Glucose Oxidase; C021538:L-mandelate dehydrogenase; C021497:lactate 2-monooxygenase