Myristic acid binding to human serum albumin investigated by dialytic exchange rate.
Dialysis rate determinations of several fatty acids in the absence of albumin revealed that the myristate anion, like that of laurate, in aqueous solution, pH 7.5, is present as a monomer anion when the concentration is below 25 microM. Palmitate and oleate solutions, on the other hand, show a tendency to aggregation even at concentrations below 0.5 microM. Multiple binding of myristate to human serum albumin in phosphate buffer, at pH 7.5, 37 degrees C, was investigated by exchange of 14C-labeled myristate across a dialysis membrane under conditions of binding equilibrium. A binding isotherm was established by least squares fitting of the stoichiometric binding constants in the stepwise binding equation to the experimental data. The best-fit solution was supplemented with 30 acceptable solutions within a probability limit of 0.95. A concept of one or two distinct high-affinity sites for binding of fatty acids could not be verified; the observations allow a variety of binding mechanisms ranging from cooperativity of the first two myristates to a model with four equal and independent sites.
Pedersen
A O
AO
Institute of Medical Biochemistry, University of Aarhus, Denmark.
Brodersen
R
R
D003956:Dialysis / Q000379:methods; D006801:Humans; D007700:Kinetics; D007850:Lauric Acids / Q000378:metabolism; D008962:Models, Theoretical; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism*; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000378:metabolism; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*
D007850:Lauric Acids; D009227:Myristic Acids; D010169:Palmitic Acids; D012709:Serum Albumin; D019814:Myristic Acid; C030358:lauric acid; D019308:Palmitic Acid