title: Graphical analysis of binding data reflecting competition between two ligands for the same acceptor sites.
abstract: A theoretical expression is derived for the analysis of results from competitive binding studies in which two multivalent ligands compete for acceptor sites, and a linear transform is suggested for simple graphical representation and assessment of experimental results. The protocol is illustrated by application to competitive binding data, obtained by ultrafiltration, on the interactions of bovine serum albumin with two structurally similar organic anions, methyl orange and methyl red. In a second experimental study the present approach is then used to establish that lactate dehydrogenase and aldolase compete for the same myofibrillar sites of bovine cardiac muscle. Finally, numerically simulated behavior of systems with additional binding sites for either ligand is used to emphasize that the criterion for classical (complete) competition is agreement between an experimentally determined equilibrium constant for ligand binding and the apparent value deduced from competitive binding studies. Nevertheless, the present analysis of competitive binding data should still offer considerable scope for screening quantitatively the cross-reactivities of drug and antigen analogs for their respective specific protein-acceptor sites.
authors: Department of Biochemistry, University of Queensland, St. Lucia, Australia.
authors: S J
authors: SJ
authors: Harris
authors: D J
authors: DJ
authors: Winzor
mesh_terms: D000818:Animals; D001391:Azo Compounds / Q000378:metabolism; D001665:Binding Sites; D001667:Binding, Competitive; D002417:Cattle; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008024:Ligands; D008956:Models, Chemical; D009132:Muscles / Q000201:enzymology; D012710:Serum Albumin, Bovine / Q000378:metabolism; D014462:Ultrafiltration
chemical_list: D001391:Azo Compounds; D008024:Ligands; D012710:Serum Albumin, Bovine; C008492:methyl red; C100258:methyl orange; D007770:L-Lactate Dehydrogenase; D005634:Fructose-Bisphosphate Aldolase