Comparative studies of the binding of some ligands to human serum albumin non-covalently attached to immobilized Cibacron Blue, or covalently immobilized on Sepharose, by column affinity chromatography.
A comparative study of the ligand-binding properties of human serum albumin was performed by the technique of affinity chromatography with the protein attached to immobilized Cibacron Blue F3GA (Blue Sepharose), or covalently immobilized on Sepharose. The binding strength of octanoate, decanoate and dodecanoate is much weaker when human serum albumin is attached to immobilized Cibacron Blue, indicating that the binding sites for fatty acids are involved in the attachment of human serum albumin to immobilized Cibacron Blue. The results revealed additional alterations of the ligand binding when human serum albumin was attached to immobilized Cibacron Blue, involving sites outside of the binding domains of fatty acids. Thus the stereoselective binding of L-tryptophan was abolished, and the resolution of the warfarin enantiomers was impaired. However, the binding strength of warfarin and salicylic acid was rather close to the values observed with human serum albumin covalently immobilized on Sepharose. It is suggested that the availability of the binding sites for L-tryptophan, warfarin and salicylic acid is partially blocked by the complex between albumin and the dye without direct participation in the complex-formation. An alternative interpretation involves an allosteric mechanism brought about by complex-formation between serum albumin and the immobilized Cibacron Blue.
D000873:Anthracenes; D001665:Binding Sites; D002846:Chromatography, Affinity / Q000379:methods; D005227:Fatty Acids / Q000378:metabolism; D006801:Humans; D008024:Ligands; D011134:Polysaccharides; D012459:Salicylates / Q000378:metabolism; D012685:Sepharose / Q000031:analogs & derivatives; D012709:Serum Albumin / Q000378:metabolism*; D014364:Tryptophan / Q000378:metabolism; D014859:Warfarin / Q000378:metabolism
D000873:Anthracenes; D005227:Fatty Acids; D008024:Ligands; D011134:Polysaccharides; D012459:Salicylates; D012709:Serum Albumin; C017842:cibacron blue-sepharose; D014859:Warfarin; D014364:Tryptophan; D012685:Sepharose