Capillary electrophoresis frontal analysis for the study of flavonoid interactions with human serum albumin.
Capillary electrophoresis based on the principles of frontal analysis (CE-FA) was used to characterize the binding of flavonoids to human serum albumin (HSA) at near-physiological conditions: 67 mM phosphate buffer (pH 7.4), temperature 36.5 degrees C. The studied flavonoids (flavone, rutin, quercitrin) displayed moderate affinities toward the human serum albumin with binding constants in the range 10(3)-10(4) M(-1). The binding of the flavonoids to the protein noticeably depended on their lipophilicity and decreased in the case of glycosylation. The corresponding thermodynamic parameters characterized the acting forces between the HSA and flavonoids as mainly hydrophobic forces and electrostatic interactions. Based on the results of the displacement experiments, the binding of the flavonoids took place at site I of the HSA molecule. The results demonstrated by CE-FA were similar to those obtained by fluorescence spectroscopy. The developed method proved to be a reliable alternative to conventional methods, providing a lot of useful parameters for characterization of ligand-protein interactions.
Knjazeva
Tatjana
T
Department of Chemistry, Faculty of Science, Tallinn University of Technology, Akadeemia tee 15, 12618 Tallinn, Estonia. tatjana.knjazeva@yahoo.com
Kaljurand
Mihkel
M
D001665:Binding Sites; D019075:Electrophoresis, Capillary / Q000379:methods*; D005419:Flavonoids / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D011485:Protein Binding; D012709:Serum Albumin / Q000737:chemistry*; D055672:Static Electricity; D013816:Thermodynamics
D005419:Flavonoids; D012709:Serum Albumin