Binding of lithocholate and its glucuronide and sulfate by human serum albumin.

In the present study, the binding affinities of lithocholate sulfate and glucuronide by human serum albumin were compared to that of lithocholate by equilibrium dialysis, and the binding sites of these bile acids on those of various fluorescent probes and bilirubin were also studied. The dissociation constants for the primary binding sites for lithocholate sulfate and glucuronide on human serum albumin were 0.057 and 0.24 microM, respectively, which were lower than that for lithocholate (0.82 microM). Lithocholate sulfate and glucuronide, as well as lithocholate, did not simply inhibit the binding of the site II and III fluorescent probes or bilirubin to albumin. Inhibition by these bile acids of the site I fluorescent probe binding to albumin suggested that the secondary binding sites of these bile acids are equal to site I. The results of simultaneous equilibrium dialysis using [3H]lithocholate and [14C]lithocholate sulfate indicated that these compounds have the same primary binding site. In conclusion, sulfation and glucuronidation may increase the binding affinities of bile acids to human serum albumin without changing their binding site.