Antigenic determinant and interspecies cross-reactivity of a monoclonal antibody to poly(ADP-ribose) synthetase.
A monoclonal antibody (1F4) was prepared against calf thymus poly(ADP-ribose) synthetase. It was classified as IgG1/kappa and its antigenic determinant was localized on the 46 kDa portion of the enzyme molecule which contains the site for the binding of DNA. When calf thymus DNA-binding proteins were subjected to immunostaining after electrophoresis and transblotting to a nitrocellulose filter, the native enzyme (120 kDa) and its endogenous degradation products (80, 64 and 32 kDa) were detected. When the interspecies cross-reactivity was examined using DNA-binding proteins from 6 different sources, 1F4 reacted with the 120- and 32-kDa protein bands in HeLa cells, mouse testis and chicken liver as in the case of calf thymus. These results indicate that the antigenic structures of poly(ADP-ribose) synthetase and its degradation products are highly conserved in various animal cells.
Kameshita
I
I
Yamamoto
H
H
Fujimoto
S
S
Shizuta
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002417:Cattle; D002645:Chickens; D000939:Epitopes / Q000276:immunology*; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D008099:Liver / Q000201:enzymology; D008297:Male; D051379:Mice; D009244:NAD+ Nucleosidase / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D011065:Poly(ADP-ribose) Polymerases / Q000276:immunology*; D013045:Species Specificity; D013737:Testis / Q000276:immunology; D013950:Thymus Gland / Q000201:enzymology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010446:Peptide Fragments; D011065:Poly(ADP-ribose) Polymerases; D009244:NAD+ Nucleosidase