Chromatography of Trypsin and Its Derivatives

Equilibrium chromatography of trypsinogen and trypsin was carried out on sulfoethyl Sephadex in tris(hydroxymethyl)aminomethane chloride buffer under conditions which limit autodigestion. At least five active forms of trypsin could be resolved. In addition to the single chain form, a second active form was shown to arise during the activation process and to be abundant in all samples. This new form was found to contain an intrachain split between lysine-131 and serine-132, and to differ in esterase and amidase behavior from the classical, single chain form of trypsin.