Chiral complexation and aggregation of bilirubin with serum albumin at a liquid/liquid interface

摘要

The chiral complexation of bilirubin (BR) with bovine and human serum albumin (BSA and HSA), and the aggregation of the complexes at the heptane+chloroform(5:1)/water interface were studied via UV/Vis absorption and circular dichroism (CD) measurements in combination with the centrifugal liquid membrane (CLM) method. The interfacial adsorptivities of BR, BSA and their complexes were also studied by performing interfacial tension measurements at the interface. The changes in the absorbances and the induced CD amplitudes of the interfacial BR–BSA complex provided insights into the mechanism of the conformational enantioselective complexation at the interface, and indicated that the chiral conversion induced by the complexation with BSA was from the P(+) form to the M(?) form of BR. The broadening of the 450 nm band and the appearance of a new shoulder at 474 nm further supported the formation of aggregates of the complexes at the interface. The dependence of the CD amplitude on the molar ratio of BSA to BR revealed that the composition of the complex was 1:1 BSA:BR. The probable interfacial reaction scheme was proposed, and the affinity constant of BR–BSA at the interface was found to be 4.67 × 108 M?2. The interfacial complexation and aggregation of BR and HSA were weaker than those of the BR–BSA complex due to the different BR binding positions adopted for BSA and HSA and the binding effect of chloroform.