Fluorometric probing on the binding of hematoxylin to serum albumin.
The binding of a cell nucleus stain, hematoxylin (HTL), to bovine serum albumin (BSA) was studied by spectroscopy including fluorescence spectra, UV-Visible absorption, circular dichroism (CD) spectra, synchronous and three-dimensional fluorescence spectra. The results indicated that the binding had led to static fluorescence quenching, with non-radiation energy transfer happening within single molecule. The observed binding constant was calculated to be 10(5.588) l mol(-1) at 311 K and one binding site had formed. The thermodynamic parameters of the interaction complied with DeltaG (theta) < 0, DeltaH (theta) < 0, DeltaS (theta) < 0 and the results indicate that hydrogen bonds played major role in the reaction. The distance r between donor (BSA) and acceptor (HTL) was obtained according to the Förster theory of non-radiation energy transfer. The structural change of BSA molecules with addition of HTL was analyzed and the optimized geometry of HTL-BSA was investigated by fluorescence probe method.
Zhang
Hua-Xin
HX
College of Chemical and Pharmaceutical Engineering, Jingchu University of Technology, 448000 Jingmen, Hubei, People's Republic of China. h.x.zhang@yeah.net
Gao
Song
S
Xiong
Ze-Yun
ZY
Liu
Shan-Pei
SP
D006416:Hematoxylin / Q000737:chemistry / Q000378:metabolism*; D007052:Ibuprofen / Q000737:chemistry; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013816:Thermodynamics; D014859:Warfarin / Q000737:chemistry
D012710:Serum Albumin, Bovine; D014859:Warfarin; D007052:Ibuprofen; D006416:Hematoxylin