Analysis of the catalytic domain of phosphotransferase activity of two avian sarcoma virus-transforming proteins.

Proteolytic digestion of the transforming protein of Rous sarcoma virus (pp60src) with trypsin, chymotrypsin, or thermolysin generated a 29,000-dalton fragment representing the carboxyl half of this molecule. This proteolytic fragment was able to phosphorylate pp60src-specific immunoglobulin as well as exogenous substrates such as angiotensin, casein, and tubulin. When quantitated on a molar basis, the protease-resistant fragment of pp60src had a greater specific activity than the intact enzyme. Digestion of pp90yes, the transforming protein of Y73 sarcoma virus with these proteases yielded a peptide of similar molecular weight which was capable of autophosphorylation as well as the phosphorylation of exogenous substrates. The proteolytic fragments of both pp60src and pp90yes displayed the same strict specificity for phosphorylation of tyrosine as the intact enzymes. These results indicate that the 29,000-dalton carboxyl end of pp60src and pp90yes can function independently as phosphotransferases and indicate that the catalytic domains of these molecules have a conformation which confers protection against limited conditions of proteolysis.