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.
MeSH Terms
Amino Acid Sequence
Animals
Carrier Proteins/genetics,metabolism
Cloning, Molecular
Haplorhini
Humans
Kidney/enzymology
Liver/enzymology
Maltose-Binding Proteins
Molecular Sequence Data
Oxidoreductases Acting on CH-NH Group Donors/genetics,isolation & purification,metabolism
Oxidoreductases, N-Demethylating/chemistry,metabolism
Peroxisomal Disorders/metabolism
Peroxisome-Targeting Signal 1 Receptor
Peroxisomes/metabolism
Phylogeny
Pipecolic Acids/blood,metabolism
Receptors, Cytoplasmic and Nuclear/metabolism
Recombinant Proteins/genetics,metabolism
Sarcosine/blood
Sarcosine Oxidase
Sequence Analysis, Protein
Sequence Homology, Amino Acid
Substrate Specificity
Chemicals
Carrier Proteins
Maltose-Binding Proteins
Peroxisome-Targeting Signal 1 Receptor
Pipecolic Acids
Receptors, Cytoplasmic and Nuclear
Recombinant Proteins
Oxidoreductases Acting on CH-NH Group Donors
Oxidoreductases, N-Demethylating
PIPOX protein, human
Sarcosine Oxidase
L-pipecolate dehydrogenase
pipecolic acid
Sarcosine
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Dodt G
Institut für Physiologische Chemie, Ruhr-Universität Bochum, 44780 Bochum, Federal Republic of Germany.
Kim D G
Reimann S A
Reuber B E
McCabe K
Gould S J
Mihalik S J
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