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Identification and expression of a new splicing variant of FAD-sulfhydryl oxidase in adult rat brain.

By Jean Radom, Didier Colin, Franck Thiebault, Mai Dognin-Bergeret, Georges Mairet-Coello, Annick Esnard-Feve, Dominique Fellmann and Michèle Jouvenot

Abstract

International audienceFlavoproteins of the quiescin/sulfhydryl oxidase (QSOX) family catalyze oxidation of peptide and protein thiols to disulfides with the reduction of oxygen to hydrogen peroxide. We report here the molecular cloning of a new putative sulfhydryl oxidase cDNA, rQSOX-L (GenBank Accession no ), from adult rat brain and its expression studied by RT-PCR, Northern and Western blots in rat tissues. DNA-sequencing demonstrated the existence of two cDNAs in rat cortex, corresponding to a long transcript (rQSOX-L) and a short transcript (rQSOX-S) which differed by 851 nucleotides due to alternative splicing. The new transcript, rQSOX-L (3356 nucleotides), was specifically expressed in brain, hypophysis, heart, testis and seminal vesicle. The distribution of this variant is not homogeneous in the different tissues studied and suggests a complex gene regulation. The full-length rQSOX-L cDNA has an open reading frame of 2250-bp encoding a protein of 750 amino acids that contains a signal peptide sequence, a protein-disulfide-isomerase-type thioredoxin and ERV1-ALR domains and a long form specific C-terminal extension. The rQSOX-L protein is highly homologous to members of the sulfhydryl oxidase/Quiescin family and contains particularly two potential sites for N-glycosylation. This protein isoform was specifically detected in rat brain tissues in opposition to the low molecular form that was ubiquitous. Matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis of the immunoprecipitate tryptic fragments allowed the identification of rQSOX-L protein

Topics: MESH: Alternative Splicing, MESH: Amino Acid Sequence, MESH: Male, MESH: Molecular Sequence Data, MESH: Oxidoreductases, MESH: RNA, Messenger, MESH: Rats, MESH: Rats, Sprague-Dawley, MESH: Transcription, Genetic, MESH: Animals, MESH: Brain, MESH: Cerebral Cortex, MESH: Flavin-Adenine Dinucleotide, MESH: Gene Expression, MESH: Genome, MESH: Glycosylation, MESH: Immunoprecipitation, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology
Publisher: 'Elsevier BV'
Year: 2006
DOI identifier: 10.1016/j.bbaexp.2006.04.008
OAI identifier: oai:HAL:hal-00482217v1
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