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Binding of the Bioactive Component Daphnetin to Human Serum Albumin Demonstrated Using Tryptophan Fluorescence Quenching
Author(s) -
Liu Jiaqin,
Tian Jianniao,
Li Ying,
Yao Xiaojun,
Hu Zhide,
Chen Xingguo
Publication year - 2004
Publication title -
macromolecular bioscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.924
H-Index - 105
eISSN - 1616-5195
pISSN - 1616-5187
DOI - 10.1002/mabi.200300109
Subject(s) - chemistry , human serum albumin , quenching (fluorescence) , tryptophan , binding constant , fluorescence , fluorescence spectroscopy , protein secondary structure , enthalpy , circular dichroism , crystallography , analytical chemistry (journal) , binding site , organic chemistry , biochemistry , amino acid , physics , quantum mechanics
Abstract Summary: Daphnetin (7,8‐dihydroxycoumarin), one of the major bioactive components isolated from Daphne koreane Nakai, has been used in traditional Chinese medicine for the treatment of coagulation disorders. It is also a chelator, an antioxidant and a protein kinase inhibitor. In this paper, a combination of intrinsic fluorescence, Fourier transform infrared (FT‐IR) spectroscopy and circular dichroic (CD) spectroscopy has been used to characterize the binding between daphnetin and human serum albumin (HSA) under physiological conditions with drug concentrations of 6.7 × 10 −6 − 2.3 × 10 −5 mol · L −1 , and a HSA concentration of 1.5 × 10 −6 mol · L −1 . Changes in the CD spectra and FT‐IR spectra were observed upon ligand binding, and the degree of tryptophan fluorescence quenching did change 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 (Δ H ) and the entropy change (Δ S ) were calculated to be −12.45 kJ · mol −1 and 52.48 J · mol −1 · K ‐1 according to the van't Hoff equation, which indicated that hydrophobic and electrostatic interactions played the main role in the binding of daphnetin to HSA, in accordance with the results of calculations performed on a Silicon Graphics Ocatane2 workstation. In addition, the binding distance between daphnetin and HSA was obtained (4.02 nm) based on the Förster energy transfer theory.The binding mode between daphnetin and HSA.