Spectroscopic and molecular docking techniques study of the interaction between oxymetholone and human serum albumin

In this study, the binding of oxymetholone (OXM), a doping drug, to human serum albumin (HSA) was explored at pH 7.40 by spectroscopic methods including spectrofluorimetry, three dimensional excitation-emission matrix (3D EEM), UV-vis absorption, resonance rayleigh scattering (RRS) and molecular docking. The fluorescence results showed that there was a considerable quenching of the intrinsic fluorescence of HSA upon binding to OXM by static quenching mechanism. The Stern-Volmer quenching constants (K-SV) between OXM and HSA at three different temperatures 295, 303, 308 K, were obtained as 4.63 x 10(4), 3.05 x 10(4) and 1.49 x 10(4) L mol(-1), respectively. Furthermore this interaction was confirmed by UV-vis spectrophotometric and RRS techniques. The binding site number, n, apparent binding constant, K-b, and corresponding thermodynamic parameters (Delta S, Delta H and Delta G) were measured at different temperatures. The Van der Waals and hydrogen-bond forces were found to stabilize OXM-HSA complex. The distance (r) between the donor and acceptor was obtained from Forster's theory of fluorescence resonance energy transfer (FRET) and found to be 1.67 nm. The 3D EEM showed that OXM slightly changes the secondary structure of HSA. Furthermore, the molecular docking was employed for identification of drug binding sites and interaction of OXM with amino acid residues. (C) 2014 Elsevier B.V. All rights reserved.