Spectroscopic probes of the individual and combined effects of Triton X-100 and chloroform on serum albumins and serum-albumin.bilirubin complexes.
The effects of the non-ionic surfactant Triton X-100 on the biphasic induced CD spectra of bilirubin complexes of human and bovine serum albumins (HSA and BSA) are divergent. While Triton X-100 inverts the induced CD spectrum of HSA.bilirubin, this surfactant enhances the ellipticity values of induced CD of BSA.bilirubin without inversion. The effect of Triton X-100 on the characteristic ultraviolet-CD spectra of the albumins are similar; both the albumins are denatured from their native globular structures. The anionic surfactant SDS, unlike non-ionic Triton X-100, dislodges the ligand from its protein complexes, indicating that both electrostatic and hydrophobic forces are involved in binding of bilirubin to the albumins. The aprotic solvent chloroform inverts the biphasic induced CD spectra of HSA.bilirubin and BSA.bilirubin, whereas CHCl3 has relatively little effect on the ultraviolet CD spectra of the albumins. The combined effect of Triton X-100 and CHCl3 shows that the effect of CHCl3 predominates over that of Triton X-100. The perturbing effects of Triton X-100 and CHCl3 on the CD or induced CD spectra of the proteins or their bilirubin complexes are reversible, and independent of the order in which components were added. The observations suggest that the denaturation of the albumins by Triton X-100 or solvation of CHCl3 within albumins markedly alter the internal topography or dynamics of the receptor sites, triggering alterations of the chirality of the bound pigment in sign and/or magnitude.
Patra
S K
SK
Department of Biochemistry and Biophysics, University of Kalyani, India.
Pal
M K
MK
D000818:Animals; D001663:Bilirubin / Q000378:metabolism*; D002417:Cattle; D002725:Chloroform / Q000494:pharmacology*; D002942:Circular Dichroism; D003902:Detergents / Q000494:pharmacology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D046911:Macromolecular Substances; D017830:Octoxynol / Q000494:pharmacology*; D011485:Protein Binding; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation; D012709:Serum Albumin / Q000187:drug effects / Q000378:metabolism*; D012997:Solvents / Q000494:pharmacology*; D013054:Spectrophotometry, Atomic
D003902:Detergents; D046911:Macromolecular Substances; D012709:Serum Albumin; D012997:Solvents; D002725:Chloroform; D017830:Octoxynol; D001663:Bilirubin