[Spectroscopic study on interaction of rodenticide brodifacoum with bovine serum albumin].
The mutual interaction of bovine serum albumin (BSA) with brodifacoum (3-[3-(4'-bromophenyl-4) 1,2,3,4-tetralin-10]-4-hydroxyl-coumarin), an anticoagulant rodenticide, was investigated by ultra-violet spectroscopy, flurorescence spectroscopy and synchronous fluorescence spectroscopy under physiological conditions. It was proved that the intrinsic fluorescence quenching of BSA by brodifacoum was the result of the formation of brodifacoum-BSA complex. And this quenching is mainly due to static fluorescence quenching. The quenching rate constant (K(sv)), binding site number (n) and binding constant (KA) at different temperatures were calculated from the double reciprocal Lineweaver-Burk plots and the quenching function of lg[(F0 - F)/F] - lg[Q] plots. The thermodynamic parameters indicated that the process of binding was a spontaneous molecular interaction and the hydrophobic force played a major role in stabilizing the brodifacoum BSA complex. The binding distance r between brodifacoum and BSA was 2.84 and 2.87 nm at 20 and 30 degrees C, respectively, which was obtained based on Forster theory of non-radiation energy transfer. The synchronous spectroscopy of BSA and brodifacoum-BSA revealed that the BSA conformation had changed in the presence of brodifacoum. The binding mode and interaction mechanism were suggested as follows: brodifacoum molecules are closed with amino acid residues with electric charge on the hydrophobic cavities of BSA by electrostatic interaction, and binded to the Trp212 residues inside of BSA hydrophobic cavities by hydrophobic interaction force, thereby changed the microenvironment around the Trp residues. The interaction prevented the energy transfer between Tyr and Trp residues, moreover, caused to a non-radiation energy transfer from Trp residues in BSA to brodifacoum, and finally leaded of the quenching the intrinsic fluorescence of BSA.
Duan
Yun-Qing
YQ
College of Art and Science, Shanxi Agricultural University, Taigu 030801, China. dddy56@sina.com
Lei
Huan-Gui
HG
Min
Shun-Geng
SG
Duan
Zhi-Qing
ZQ
D015110:4-Hydroxycoumarins / Q000737:chemistry*; D001665:Binding Sites; D004735:Energy Transfer; D057927:Hydrophobic and Hydrophilic Interactions; D012378:Rodenticides / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
D015110:4-Hydroxycoumarins; D012378:Rodenticides; D012710:Serum Albumin, Bovine; C013418:bromfenacoum