cDNA cloning, gene organization and expression analysis of human peptidylarginine deiminase type I.
Peptidylarginine deiminases (PADs) catalyse a post-translational modification of proteins through the conversion of arginine residues into citrullines. The existence of four isoforms of PAD (types I, II, III and IV) encoded by four different genes, which are distinct in their substrate specificities and tissue-specific expression, was reported in rodents. In the present study, starting from epidermis polyadenylated RNA, we cloned by reverse transcriptase-PCR a full-length cDNA encoding human PAD type I. The cDNA was 2711 bp in length and encoded a 663-amino-acid sequence. The predicted protein shares 75% identity with the rat PAD type I sequence, but displays only 50-57% identity with the three other known human isoforms. We have described the organization of the human PAD type I gene on chromosome 1p36. A recombinant PAD type I was produced in Escherichia coli and shown to be enzymically active. Human PAD type I mRNAs were detected by reverse transcriptase-PCR not only in the epidermis, but also in various organs, including prostate, testis, placenta, spleen and thymus. In human epidermis extracts analysed by Western blotting, PAD type I was detected as a 70 kDa polypeptide, in agreement with its predicted molecular mass. As shown by immunohistochemistry, the enzyme was expressed in all the living layers of human epidermis, with the labelling being increased in the granular layer. This is the first description of the human PAD type I gene and the first demonstration of its expression in epidermis.
Guerrin
Marina
M
Department of Epidermal Differentiation and Rheumatoid Autoimmunity, Toulouse-Purpan Pathophysiology Center, INSERM U563 - P. Sabatier University (IFR30, INSERM-CNRS-P. Sabatier Université-Centre Hospitalier Universitaire), Toulouse, France.
Ishigami
Akihito
A
Méchin
Marie-Claire
MC
Nachat
Rachida
R
Valmary
Séverine
S
Sebbag
Mireille
M
Simon
Michel
M
Senshu
Tatsuo
T
Serre
Guy
G
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004817:Epidermis / Q000201:enzymology; D006801:Humans; D006867:Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D000080062:Protein-Arginine Deiminase Type 1; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D018076:DNA, Complementary; D012333:RNA, Messenger; D011994:Recombinant Proteins; D006867:Hydrolases; D000080062:Protein-Arginine Deiminase Type 1