Assessment of the europium(III) binding sites on albumin using fluorescence spectroscopy

J Phys Chem B. 2014 Jun 19;118(24):6626-33. doi: 10.1021/jp501277z. Epub 2014 Jun 10.

Abstract

Intrinsic fluorescence quenching of bovine serum albumin (BSA) and europium(III) luminescence in BSA complexes were investigated. The number of BSA binding sites (n) and equilibrium constant (Keq) values were determined from both measurements provided qualitatively different results. While the modified Stern-Volmer relation for BSA fluorescence quenching gave n = 1 at pH 4.5 and pH 6, two sets of binding sites were determined from Eu(3+) luminescence with n1 = 2, n2 = 4 at pH 6 and n1 = 1, n2 = 2 at pH 4.5. The model explaining the discrepancy between the results obtained by these fluorescent approaches was suggested, and the limitations in application of the "log-log" Stern-Volmer plots in analysis of binding processes were discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Cattle
  • Europium / chemistry*
  • Europium / metabolism
  • Hydrogen-Ion Concentration
  • Kinetics
  • Protein Binding
  • Serum Albumin, Bovine / chemistry*
  • Serum Albumin, Bovine / metabolism
  • Spectrometry, Fluorescence

Substances

  • Serum Albumin, Bovine
  • Europium