Palmitate and stearate binding to human serum albumin. Determination of relative binding constants.
Multiple binding equilibria of two apparently insoluble ligands, palmitate and stearate, to defatted human serum albumin were studied in a 66 mM sodium phosphate buffer (pH 7.4) at 37 degrees C, by determination of dialytic exchange rates of ligands among identical equilibrium solutions. The experimental data were analysed by a computerised curve fitting procedure using equilibrium equations for multiple binding of ligands, containing relative binding constants, valid whether the ligands are truly insoluble or are slightly soluble and irrespective of aggregation in aqueous solution. A best-fit set of relative binding constants was found, and subsequently 30 sets of acceptable constants for each set of data in order to evaluate the variation. The data were first fitted by the relative Scatchard's equation, then by the relative, stoichiometric equation. Scatchard's equation is deduced on the presumption that cooperativity is absent while the stoichiometric equation is valid even when cooperativity is present. It was found with palmitate as well as with stearate that the two equations fitted the data equally well, and it was concluded that the observations were compatible with absence of cooperativity. The relative Scatchard binding constants were converted to relative, stoichiometric constants and it was found that the variations of the latter were slight.
Vorum
H
H
Department of Medical Biochemistry, University of Aarhus, Denmark. hv@biokemi.aau.dk
Fisker
K
K
Honoré
B
B
D006801:Humans; D007700:Kinetics; D008956:Models, Chemical; D019308:Palmitic Acid / Q000097:blood*; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*; D012995:Solubility; D013229:Stearic Acids / Q000097:blood*
D012709:Serum Albumin; D013229:Stearic Acids; D019308:Palmitic Acid; C031183:stearic acid