DISC ELECTROPHORESIS OF IRRADIATED TRYPSIN*

Abstract

Abstract— Trypsin inactivated by u.v. radiation, gamma radiation, visible radiation in the presence of sensitizing dyes, and autolysis, was examined by the method of disc electrophoresis. Untreated Worthington twice crystallized salt‐free trypsin separated into four bands which moved toward the cathode; the main band, which had the greatest mobility, contained all of the detectable tryptic activity. The next most mobile band has been assumed to be a chymotrypsin contaminant. The other two bands are of unknown nature. A progressive loss of all the bands was observed when the enzyme was inactivated by those procedures which produce a ‘damaged’ class of trypsin molecules, i.e. flavin‐sensitized photooxidation, autolysis, and treatment with u.v. and gamma radiation. No loss of the main band was observed during photoinactivation with methylene blue and eosin Y as sensitizers. In this latter case, it is postulated that the trypsin inactivation products must be of such a nature that the net charge and conformation of the protein is not greatly changed, thus permitting all of the protein to remain in the same band during electrophoresis.