Bilirubin, model membranes and serum albumin interaction: The influence of fatty acids.
Electronic circular dichroism (ECD), absorption and fluorescence spectroscopy were used to study the enantioselective interactions which involved bilirubin (BR), liposomes, human serum albumin of two different purities, pure (HSA) and non-purified of fatty acids (FA-HSA), and individual fatty acids. The application of the ECD technique to such a complex problem provided a new perspective on the BR binding to liposomes. Our results demonstrated that in the presence of pure HSA, BR preferred to bind to the protein over the liposomes. However, in the presence of FA-HSA, BR significantly bound to the liposomes composed either of DMPC or of sphingomyelin and bound only moderately to the primary and secondary binding sites of FA-HSA even at high BR concentrations. For the DMPC liposomes, even a change of BR conformation upon binding to the primary binding site was observed. The individual saturated fatty acids influenced the BR binding to HSA and liposomes in a similar way as fatty acids from FA-HSA. The unsaturated fatty acids interacted with BR alone and prevented it from interacting with either 99-HSA or the liposomes. In the presence of arachidonic acid, BR interacted enantioselectively with the liposomes and only moderately with 99-HSA. Hence, our results show a substantial impact of the liposomes on the BR binding to HSA. As a consequence of the existence of fatty acids in the blood plasma and in the natural structure of HSA, BR may possibly bind to the cell membranes even though it is normally bound to HSA.
Department of Analytical Chemistry, University of Chemistry and Technology, Prague, Technická 5, 166 28 Prague 6, Czech Republic.
Pavlína
P
Novotná
Department of Physics and Measurements, University of Chemistry and Technology, Prague, Technická 5, 166 28, Prague 6, Czech Republic. Electronic address: Marie.Urbanova@vscht.cz.
Marie
M
Urbanová
D001663:Bilirubin / Q000378:metabolism*; D002942:Circular Dichroism; D005227:Fatty Acids / Q000378:metabolism*; D006801:Humans; D008081:Liposomes / Q000737:chemistry; D008567:Membranes, Artificial; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D005227:Fatty Acids; D008081:Liposomes; D008567:Membranes, Artificial; D012709:Serum Albumin; D001663:Bilirubin
Bilirubin–albumin interaction; Bilirubin–lipid interaction; Electronic circular dichroism; Fatty acid; Liposome