title: Trypsin and affinity chromatography.
abstract: Affinity adsorbents for trypsin which were prepared by immobilizing product-type ligands, that is, peptides having C-terminal arginine, proved to be effective not only for preparative purposes but also for basic research on molecular recognition. The properties of the binding site of trypsin were revealed by chromatographic experiments. Quantitative analysis based on the theory of frontal affinity chromatography proved to be extremely effective. As an extension of the product-type ligands, peptide argininals were also used and information on the mechanism of action of these inhibitors was obtained. Anhydrotrypsin, which lost the hydroxyl group of Ser183, was found to gain increased binding ability for product-type compounds. This inactivated enzyme was also used as an immobilized ligand and the unique affinity adsorbent thus prepared proved to be extremely effective for the separation of peptides and recombinant proteins based on their C-terminal structures. High-performance affinity chromatography of trypsin and related enzymes using a polymer-based support was also developed.
authors.affiliation: Department of Biochemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Kanagawa, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Kasai
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity; D004800:Enzymes, Immobilized; D006801:Humans; D011485:Protein Binding; D014357:Trypsin / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D004800:Enzymes, Immobilized; D014357:Trypsin