A mammalian tryptophanyl-tRNA synthetase is associated with protein kinase activity

Bovine Trp-tRNA synthetase is a dimer with subunit molecular mass of 60 kDa (p60) which catalyzes ATP-dependent formation of tryptophanyl-tRNA. Evidence is presented that Trp-tRNA synthetase whose homogeneity had been proven by SDS/PAGE and silver staining of the gel is autophosphorylated in vitro. Anti-(Trp-tRNA synthetase) antibodies, whose specificity was verified by using a combination of different approaches, were able to effectively inhibit and immunoprecipitate the Trp-tRNA-synthetase-associated kinase activity. The two-dimensional tryptic phosphopeptide map of autophosphorylated p60 Trp-tRNA synthetase was found to be similar to that of its major 40-kDa degradation fragment bearing resemblance to previously demonstrated unlabeled peptide patterns of the Trp-tRNA synthetase forms. Trp-tRNA synthetase which had undergone denaturation during SDS/PAGE, regained serine/threonine specific protein kinase activity (PK 60) after guanidine treatment. Trp-tRNA synthetase induced phosphorylation of specific substrate such as 100-kDa protein in non-immune but not in anti-(Trp-tRNA synthetase) sera which distinguishes Trp-tRNA-synthetase-associated kinase from other protein kinases. Sequence analysis permitted the identification of regions of bovine Trp-tRNA synthetase sharing similarity with the catalytic domains of known protein kinases. These findings suggest that PK 60 and Trp-tRNA synthetase (p60) are either closely related or identical.