Effects of ionic strength and pH on the binding of XXXX to human serum albumin.

@entity54586 @entity25014 @entity376 @entity844
Binding equilibria for the interactions of the @entity25014 anions, laurate and myristate, with defatted human serum albumin have been investigated under varying environmental conditions such as ionic strength and pH. Since these ligands bind strongly to albumin (Kass approximately 10(7) M-1), conventional equilibrium dialysis is not a feasible method for these investigations. Consequently, we employed a dialysis method, allowing determination of very low concentrations of unbound ligand by measuring the rate of exchange of labelled ligand across a dialysis membrane under conditions of chemical equilibrium. Over a range of ionic strength, 8-68 mM, the binding of the first few molecules of laurate to albumin was weakened with increasing ionic strength, whereas the binding of subsequent molecules seemed to proceed independently of ionic strength. The binding of myristate by albumin, however, appeared to be independent of ionic strength in the observed range of concentrations. The influence of pH in the range 5.1-9.0 on the binding of the two @entity844 anions by albumin was more complicated. The first molecule of @entity54586 appeared to bind with a slightly weaker affinity to albumin at low pH, compared to pH 7 and high pH, while the trends for the following molecules varied. The binding of myristate (irrespective of concentration) seemed to strengthen monotonously with pH, but this conclusion depends critically on the interpretation of the kinetic behaviour of the myristate anion. We have previously shown [Pedersen, A. O., Honor  , B. _ Brodersen, R. (1990) Eur. J. Biochem. 190, 497-502] that the strength of binding of the first few molecules of the two @entity844 anions to albumin decreases with increasing temperature, whereas binding of subsequent molecules seems to proceed independently of temperature. We explain these findings as follows. The binding of the first few (3 or 4) molecules of the C12 laurate anion is clearly driven by formation of ionic bonds between the @entity844 anion and positively charged groups, such as @entity376 residues, in the albumin molecule, whereas the binding of subsequent molecules of laurate seems to depend more on hydrophobic interactions. In the case of the C14 myristate anion, the binding of the first few (only 1 or 2) molecules may depend on ionic forces, but binding of the following molecules of myristate seems to depend on hydrophobic interactions only.(ABSTRACT TRUNCATED AT 250 WORDS)
