[Immunochemical properties of tryptophanyl-tRNA synthetase and its fragments].
The interaction between beef pancreas tryptophanyl-tRNA synthetase and its fragments produced after limited proteolysis, with IgG fraction of antiserum and with Fab fragment of IgG has been studied. Both the intact antibodies and Fab fragments inhibit the enzyme activity in tRNA aminoacylation and tryptophan dependent ATP-32P pyrophosphate exchange reactions. However, the enzyme inhibited by antibodies is still able to form a complex with tryptophanyl-tRNA. The enzymatically active fragment obtained after endogenous proteolysis interacts only with 1/3 of the antibodies against native enzyme. The fragment produced by trypsinolysis possess similar immunochemical properties. This fragment has almost the same molecular weight but is enzymatically inactive. Pure antibodies against tryptic fragment isolated by means of specific immunoabsorbent inhibit the enzymatic activity. The antibodies which do not interact with this fragment (2/3 of the total amount of antibodies) have no influence on the enzymatic activity. The immunochemical identity of the two synthetase fragments differing in their enzymatic activity supports the assumption that the loss of enzymatic activity of the tryptis fragment is caused by lack of a small peptide which is retained in case of endogenous proteolysis. Probably the amino acid residues of this peptide participate in formation of the active centre of tryptophanyl-tRNA synthetase. A new procedure for determination of the number of antigenic determinants in proteins is developed. It is shown by this method that beef pancreas tryptophanyl-tRNA synthetase contains 9 +/- 1 antigenic determinants.
D000604:Amino Acyl-tRNA Synthetases / Q000276:immunology*; D000818:Animals; D000937:Antigen-Antibody Reactions; D002417:Cattle; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D007700:Kinetics; D010179:Pancreas / Q000201:enzymology; D010446:Peptide Fragments / Q000276:immunology; D010447:Peptide Hydrolases; D014357:Trypsin; D014369:Tryptophan-tRNA Ligase / Q000276:immunology* / Q000378:metabolism
D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D010446:Peptide Fragments; D010447:Peptide Hydrolases; D014357:Trypsin; D000604:Amino Acyl-tRNA Synthetases; D014369:Tryptophan-tRNA Ligase
Immunokhimicheskie svoĭstva triptofanil-tRNK-sintetazy i ee fragmentov.