Antigenic structure of adenylate kinase from porcine skeletal muscle. IV. Two antigenic determinants on carboxyl-terminal peptide 126-194.
Delineation of the location(s) of antigenic activity in CNBr peptide 126-194 from porcine skeletal muscle adenylate kinase (AK) was attempted. Peptide 126-194 was digested with chymotrypsin, Staphylococcus aureus V8 protease and trypsin, and several short peptides were purified from the digests by reverse-phase high-performance liquid chromatography (HPLC). Inhibition of the binding of radioiodinated peptide 126-194 to goat antibody to porcine skeletal muscle AK (anti-AK antibody) by the peptides obtained by the enzymatic cleavages was examined by solid phase radioimmunoassay (RIA). At least two antigenic determinants have been identified from the results. One is in the amino (N)-terminal half region 126-154, especially in the vicinity of 131-144, and the other is in the carboxyl (C)-terminal half region 165-183, especially in the vicinity of 165-171. Both of them seem to correspond to exposed and accessible regions in the three-dimensional structure of AK. The correlation between antigenicity and high mobility of the loop in the estimated antigenic region 131-144 is also discussed.
D000263:Adenylate Kinase / Q000276:immunology*; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003488:Cyanogen Bromide; D000939:Epitopes / Q000032:analysis*; D009132:Muscles / Q000201:enzymology*; D010446:Peptide Fragments / Q000276:immunology; D010770:Phosphotransferases / Q000276:immunology*; D011487:Protein Conformation; D011863:Radioimmunoassay; D013552:Swine
D000939:Epitopes; D010446:Peptide Fragments; D010770:Phosphotransferases; D000263:Adenylate Kinase; D003488:Cyanogen Bromide