title: Affinity chromatography of serine proteases on the triazine dye ligand Cibacron Blue F3G-A.
abstract: The interaction between complement component factor B and the triazine dye ligand Cibacron Blue F3G-A coupled to a cross-linked agarose matrix (Blue Sepharose) was found to involve the Bb part of the molecule, and to be inhibited by benzamidine. Human, chicken and rainbow trout factor B which had bound to Blue Sepharose could, subsequently be eluted with benzamidine. Other serine proteases (C2, factor II, factor IX, trypsin, chymotrypsin, proteinase 3) also bound to Blue Sepharose but only those belonging to the trypsin family could be eluted with benzamidine. Trypsin treated with the active-site inhibitor phenylmethylsulfonyl fluoride did not bind to Blue Sepharose and pretreatment of Blue Sepharose with benzamidine did not influence binding of proteases. We conclude that trypsin-like serine proteases can be purified on Blue Sepharose and that the interaction of these serine proteases with Blue Sepharose involves the active site of the enzyme.
authors.affiliation: Department of Immunology, Statens Serum Institut, Copenhagen, Denmark.
authors.forename: C
authors.initials: C
authors.lastname: Koch
authors.forename: L
authors.initials: L
authors.lastname: Borg
authors.forename: K
authors.initials: K
authors.lastname: Skjødt
authors.forename: G
authors.initials: G
authors.lastname: Houen
mesh_terms: D000818:Animals; D002645:Chickens; D002846:Chromatography, Affinity / Q000379:methods*; D004396:Coloring Agents / Q000737:chemistry*; D006801:Humans; D008968:Molecular Conformation; D017686:Oncorhynchus mykiss; D012697:Serine Endopeptidases / Q000032:analysis*; D014227:Triazines / Q000737:chemistry*
chemical_list: D004396:Coloring Agents; D014227:Triazines; C012842:Cibacron Blue F 3GA; D012697:Serine Endopeptidases