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J. Biol. Chem., Vol. 276, Issue 17, 13830-13837, April 27, 2001
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,,
From Equipe "Protéases et Vectorisation," INSERM
EMI-U-0010 Université François Rabelais, Faculté de
Médecine, 2 bis Boulevard Tonnellé,
37032 Tours cedex, France
Rat FAD-dependent sulfhydryl oxidase
was purified; partial sequencing indicated that it was homologous to
human quiescin Q6. A cDNA (GenBankTM accession
no. AF285078) was cloned from rat seminal vesicles, and active
recombinant sulfhydryl oxidase was expressed in Chinese hamster ovary
epithelial cells. This 2472-nucleotide cDNA has an open reading
frame of 1710 base pairs, encoding a protein of 570 amino acids
including a 32-amino acid leader sequence and two potential sites for
N-glycosylation. One of them is used and the 64,000 Mr purified protein was transformed to 61,000 by the action of endoglycosidase F. Northern blotting and reverse transcription-polymerase chain reaction analyses showed that there were
small amounts of sulfhydryl oxidase in the rat testis, prostate, lung,
heart, kidney, spleen, and liver, and that the gene was highly
expressed in seminal vesicles and epididymis. Rat sulfhydryl oxidase
cDNA corresponds to the human cell growth inhibiting factor cDNA, which could be a differently spliced form of quiescin Q6. Comparing sulfhydryl oxidase sequences with those of human quiescin Q6
and mammalian and Caenorhabditis elegans quiescin
Q6-related genes established the existence of a new family of
FAD-dependent sulfhydryl oxidase/quiescin Q6-related genes
containing protein-disulfide isomerase-type thioredoxin and yeast ERV1 domains.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/ EBI Data Bank with accession number(s) AF285078.
These authors contributed equally to this work (A. E.-F. for
the sulfhydryl oxidase purification and characterization and B. B. for the cloning, sequencing, and expression of sulfhydryl oxidase cDNA).
§
To whom correspondence should be addressed: Laboratoire
"Enzymologie et Chimie des Protéines," INSERM EMI-U0010,
Faculté de Médecine, 2 bis Blvd. Tonnellé, 37032 Tours cedex, France. Tel.: 33-2-47-36-62-06; Fax: 33-2-47-36-60-46;
E-mail: esnard@univ-tours.fr.
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