title: Binding of genistein to human serum albumin demonstrated using tryptophan fluorescence quenching.
abstract: Genistein is an isoflavone and phytoestrogen that is a potent inhibitor of cell proliferation and angiogenesis. This study was designed to investigate the binding of genistein to human serum albumin (HSA) under physiological conditions with drug concentrations in the range of 6.7 x 10(-6) to 2.0 x 10(-5) mol L(-1) and HSA concentration at 1.5 x 10(-6) mol L(-1). Fluorescence quenching methods in combination with Fourier transform infrared (FT-IR) spectroscopy and circular dichroism (CD) spectroscopy was used to determine the binding mode, the binding constant and the protein structure changes in the presence of genistein in aqueous solution. Changes in the CD spectra and FT-IR spectra were observed upon ligand binding, and the degree of tryptophan fluorescence quenching change did significantly in the complexes. These data have proved the change in protein secondary structure accompanying ligand binding. The change in tryptophan fluorescence intensity was used to determine the binding constants. The thermodynamic parameters, the enthalpy change (DeltaH) and the entropy change (DeltaS) were calculated to be -22.24 kJ mol(-1) and 19.60 J mol(-1) K(-1) according to the van't Hoff equation, which indicated that hydrophobic and electrostatic interactions play the main role in the binding of genistein to HSA.
authors.affiliation: Mianyang Normal University, Mianyang 621000, PR China. bian-55@sohu.com
authors.forename: Qingquan
authors.initials: Q
authors.lastname: Bian
authors.forename: Jiaqin
authors.initials: J
authors.lastname: Liu
authors.forename: Jianniao
authors.initials: J
authors.lastname: Tian
authors.forename: Zhide
authors.initials: Z
authors.lastname: Hu
mesh_terms: D002942:Circular Dichroism; D005453:Fluorescence; D019833:Genistein / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011485:Protein Binding; D012709:Serum Albumin / Q000737:chemistry / Q000378:metabolism*; D012996:Solutions; D017550:Spectroscopy, Fourier Transform Infrared; D014364:Tryptophan / Q000737:chemistry*
chemical_list: D012709:Serum Albumin; D012996:Solutions; D014364:Tryptophan; D019833:Genistein