Purification of bovine liver histidyl-tRNA synthetase, the Jo-1 antigen of polymyositis: size of the whole enzyme and its characteristic proteolytic fragments.
We have recently reported the marked increase in frequency which can be achieved in the detection of the anti-Jo-1 antibody of polymyositis in serum samples by replacing commercial mixtures of cytoplasmic and nuclear antigens with the purified antigen, histidyl-tRNA synthetase. The present paper describes a method for purifying this antigen and an investigation of its size. Molecular masses previously reported for the enzyme have varied from 85-154 kDa and subunit molecular masses varying from 40-77 kDa have been observed. Several of these fragments are of sizes similar to those of a number of other autoantigens commonly observed in connective tissue diseases. Since the clinical identification of these autoantigens often relies exclusively on size determination by Western blotting, we have characterized the commonly occurring fragments of histidyl-tRNA synthetase lest they confuse such identification. It is concluded that histidyl-tRNA synthetase, like many other aminoacyl-tRNA synthetases, is subject to severe proteolysis during extraction procedures. Several characteristic fragments (Mr = 80, 75, 61, 55, 50 and 45 kDa) result, a finding that provides a satisfactory explanation of the various values previously reported. The intact bovine enzyme is a dimer of molecular mass close to 160 kDa.
D000604:Amino Acyl-tRNA Synthetases / Q000302:isolation & purification*; D000818:Animals; D000941:Antigens / Q000302:isolation & purification*; D002417:Cattle; D003593:Cytoplasm / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D006643:Histidine-tRNA Ligase / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D006868:Hydrolysis; D008970:Molecular Weight; D009220:Myositis / Q000201:enzymology / Q000276:immunology*; D010449:Peptide Mapping; D010455:Peptides / Q000032:analysis; D011233:Precipitin Tests; D014357:Trypsin
D000941:Antigens; D010455:Peptides; D014357:Trypsin; D000604:Amino Acyl-tRNA Synthetases; D006643:Histidine-tRNA Ligase