title: L-Pipecolic acid oxidase, a human enzyme essential for the degradation of L-pipecolic acid, is most similar to the monomeric sarcosine oxidases.
abstract: L-Pipecolic acid oxidase activity is deficient in patients with peroxisome biogenesis disorders (PBDs). Because its role, if any, in these disorders is unknown, we cloned the associated human gene and expressed its protein product. The cDNA was cloned with the use of a reverse genetics approach based on the amino acid sequence obtained from purified L-pipecolic acid oxidase from monkey. The complete cDNA, obtained by conventional library screening and 5' rapid amplification of cDNA ends, encompassed an open reading frame of 1170 bases, translating to a 390-residue protein. The translated protein terminated with the sequence AHL, a peroxisomal targeting signal 1. Indirect immunofluorescence studies showed that the protein product was expressed in human fibroblasts in a punctate pattern that co-localized with the peroxisomal enzyme catalase. A BLAST search with the amino acid sequence showed 31% identity and 53% similarity with Bacillus sp. NS-129 monomeric sarcosine oxidase, as well as similarity to all sarcosine oxidases and dehydrogenases. No similarity was found to the peroxisomal D-amino acid oxidases. The recombinant enzyme oxidized both L-pipecolic acid and sarcosine. However, PBD patients who lack the enzyme activity accumulate only L-pipecolic acid, suggesting that in humans in vivo, this enzyme is involved mainly in the degradation of L-pipecolic acid.
authors.affiliation: Institut für Physiologische Chemie, Ruhr-Universität Bochum, 44780 Bochum, Federal Republic of Germany.
authors.forename: G
authors.initials: G
authors.lastname: Dodt
authors.forename: D G
authors.initials: DG
authors.lastname: Kim
authors.forename: S A
authors.initials: SA
authors.lastname: Reimann
authors.forename: B E
authors.initials: BE
authors.lastname: Reuber
authors.forename: K
authors.initials: K
authors.lastname: McCabe
authors.forename: S J
authors.initials: SJ
authors.lastname: Gould
authors.forename: S J
authors.initials: SJ
authors.lastname: Mihalik
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D000882:Haplorhini; D006801:Humans; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D058619:Maltose-Binding Proteins; D008969:Molecular Sequence Data; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D010089:Oxidoreductases, N-Demethylating / Q000737:chemistry / Q000378:metabolism; D018901:Peroxisomal Disorders / Q000378:metabolism; D000074435:Peroxisome-Targeting Signal 1 Receptor; D020675:Peroxisomes / Q000378:metabolism; D010802:Phylogeny; D010875:Pipecolic Acids / Q000097:blood / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012521:Sarcosine / Q000097:blood; D050879:Sarcosine Oxidase; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D002352:Carrier Proteins; D058619:Maltose-Binding Proteins; D000074435:Peroxisome-Targeting Signal 1 Receptor; D010875:Pipecolic Acids; D018160:Receptors, Cytoplasmic and Nuclear; D011994:Recombinant Proteins; D000587:Oxidoreductases Acting on CH-NH Group Donors; D010089:Oxidoreductases, N-Demethylating; C494349:PIPOX protein, human; D050879:Sarcosine Oxidase; C032771:L-pipecolate dehydrogenase; C031345:pipecolic acid; D012521:Sarcosine