Molecular characterization of peptidylarginine deiminase in HL-60 cells induced by retinoic acid and 1alpha,25-dihydroxyvitamin D(3).
Three types of peptidylarginine deiminase (PAD), which converts a protein arginine residue to a citrulline residue, are widely distributed in animal tissues. Little is known about PAD of hemopoietic cells. We found that PAD activity in human myeloid leukemia HL-60 cells was induced with the granulocyte-inducing agents retinoic acid and dimethyl sulfoxide and with the monocyte-inducing agent 1alpha,25-dihydroxyvitamin D(3). We cloned and characterized a PAD cDNA from retinoic acid-induced cells. The cDNA was 2,238 base pairs long and encoded a 663-amino acid polypeptide. The HL-60 PAD had 50-55% amino acid sequence identities with the three known enzymes and 73% identity with the recently cloned keratinocyte PAD. The recombinant enzyme differs in kinetic properties from the known enzymes. Immunoblotting and Northern blotting with an antiserum against the enzyme and the cDNA, respectively, showed that a protein of approximately 67 kDa increased concomitantly with increase of mRNA of approximately 2.6 kilobases during granulocyte differentiation. During monocyte differentiation the same mRNA and protein increased as in granulocyte differentiation. Neither the enzyme activity nor the protein was found in macrophage-induced cells. These results suggested that expression of the PAD gene is tightly linked to myeloid differentiation.
Nakashima
K
K
Graduate School of Integrated Science, Yokohama City University, 22-2, Seto, Kanazawa-ku, Yokohama 236-0027, Japan.
Hagiwara
T
T
Ishigami
A
A
Nagata
S
S
Asaga
H
H
Kuramoto
M
M
Senshu
T
T
Yamada
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D002117:Calcitriol / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D017931:DNA Primers; D004790:Enzyme Induction; D015723:Gene Library; D006098:Granulocytes / Q000166:cytology / Q000201:enzymology; D018922:HL-60 Cells; D006801:Humans; D006867:Hydrolases / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D015603:Keratinocytes / Q000201:enzymology; D007700:Kinetics; D008969:Molecular Sequence Data; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism; D016415:Sequence Alignment; D013463:Sulfuric Acid Esters / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology*
D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D013463:Sulfuric Acid Esters; D014212:Tretinoin; D006867:Hydrolases; C000629235:PADI4 protein, human; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases; D002117:Calcitriol; C007482:dimethyl sulfate; D013755:Tetradecanoylphorbol Acetate