Rat seminal vesicle FAD-dependent sulfhydryl oxidase. Biochemical characterization and molecular cloning of a member of the new sulfhydryl oxidase/quiescin Q6 gene family.
Rat FAD-dependent sulfhydryl oxidase was purified; partial sequencing indicated that it was homologous to human quiescin Q6. A cDNA (GenBank 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 M(r) 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.
Equipe "Protéases et Vectorisation," INSERM EMI-U-0010 Université François Rabelais, Faculté de Médecine, 2 bis Boulevard Tonnellé, 37032 Tours cedex, France.
B
B
Benayoun
A
A
Esnard-Fève
S
S
Castella
Y
Y
Courty
F
F
Esnard
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000378:metabolism; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D008297:Male; D008957:Models, Genetic; D008969:Molecular Sequence Data; D005810:Multigene Family; D016366:Open Reading Frames; D010088:Oxidoreductases / Q000737:chemistry* / Q000235:genetics*; D010206:Papain / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012669:Seminal Vesicles / Q000201:enzymology*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013879:Thioredoxins / Q000737:chemistry; D013997:Time Factors; D014018:Tissue Distribution; D014162:Transfection; D050862:Oxidoreductases Acting on Sulfur Group Donors
D000596:Amino Acids; D018076:DNA, Complementary; D005182:Flavin-Adenine Dinucleotide; D010088:Oxidoreductases; D010206:Papain; D012333:RNA, Messenger; D011994:Recombinant Proteins; D013879:Thioredoxins; D050862:Oxidoreductases Acting on Sulfur Group Donors