Affinity chromatography of trypsin and related enzymes. II. An affinity adsorbent containing glycylglycyl-L-arginine.
An affinity adsorbent for trypsin [EC 3.4.21.4] (GGA Sepharose) was prepared. Glycylglycyl-L-arginine (GGA) was synthesized by a simple procedure and was immobilized on agarose gel. This adsorbent proved to have essentially the same characteristics as AP Sepharose, which is an affinity adsorbent containing tryptic peptides of protamine (1). GGS Sepharose was specific for native trypsin and had a stronger affinity at lower pH's (6-5) than at the optimum pH of trypsin action (8.2). It also proved to be suitable for analytical experiments because of its relatively weak affinity. By comparison of the elution profiles of trypsin from GGA Sepharose under various conditions, the nature of the interaction of trypsin with the adsorbent could be studied. It was found that alpha- and beta-trypsin could be distinguished. In the presence of arginine and N-substitute arginines, the elution of trypsin was accelerated. From the extents of the accelerating effects, the affinities of these compouunds could be compared.
D000818:Animals; D001120:Arginine; D001665:Binding Sites; D002417:Cattle; D002846:Chromatography, Affinity; D002918:Chymotrypsin / Q000302:isolation & purification*; D005998:Glycine; D009842:Oligopeptides; D010179:Pancreas / Q000201:enzymology; D011485:Protein Binding; D012995:Solubility; D014357:Trypsin / Q000302:isolation & purification*
D009842:Oligopeptides; D001120:Arginine; D002918:Chymotrypsin; D014357:Trypsin; D005998:Glycine