Combined biochemical and immunochemical comparison of peptidylarginine deiminases present in various tissues.
We have performed a combined biochemical and immunochemical study on the identity of peptidylarginine deiminases (EC 3.5.3.15) present in various mammalian tissues. First, we purified peptidylarginine deiminase from rat skeletal muscle. It gave a single band of molecular weight 83,000 in sodium dodecyl sulfate polyacrylamide gel electrophoresis. Next we immunized rabbits with the purified enzyme. The resulting antibodies reacted specifically with the antigen in Western blot assay. Most of the enzyme activities present in rat skeletal muscle, brain, spinal cord, submaxillary gland and spleen could be characterized as the same muscle-type enzyme by immunoprecipitation and Western blot assay. The antibodies did not react with enzyme samples obtained from rat hair follicles and bovine epidermis. The lack of immunoreactivity of the epidermal enzyme could not be accounted for by the species difference, since the antibodies reacted with a 83 kDa polypeptide of bovine brain, which was thought to represent a bovine counterpart of the muscle-type enzyme. The epidermal enzyme could be distinguished from the other enzyme samples by its high activity towards benzoylarginine. These data suggest the existence of at least three types of peptidylarginine deiminase in mammalian tissues, i.e., a muscle type, a hair follicle type, and an epidermal type.
Tokyo Metropolitan Institute of Gerontology, Japan.
K
K
Watanabe
K
K
Akiyama
K
K
Hikichi
R
R
Ohtsuka
A
A
Okuyama
T
T
Senshu
D000818:Animals; D000918:Antibody Specificity; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D004817:Epidermis / Q000201:enzymology; D005260:Female; D006197:Hair / Q000201:enzymology; D006867:Hydrolases / Q000276:immunology / Q000302:isolation & purification*; D007118:Immunoassay; D008297:Male; D009132:Muscles / Q000201:enzymology; D011233:Precipitin Tests; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases; D011817:Rabbits; D051381:Rats
D006867:Hydrolases; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases