[Studies on the action features between cefuroxime axetil and bovine serum albumin].
Under different temperatures and physiological conditions, with cefuroxime axetil concentrations in the range of 1.959 X 10(-6) to 13.71 X 10(-6) mol x L(-1), and bovine serum albumin (BSA) concentrations at 2.0 X 10(-6) mol x L(-1), the interaction between cefuroxime axetil and BSA was studied by fluorescence spectroscopy, three-dimensional fluorescence spectrum, synchronous fluorescence spectrum and UV-Vis absorption spectroscopy. After analyzing and processing the fluorescence quenching data at different temperatures according to Sterm-Volmer equation, Lineweaver-Burk equation and thermodynamic equation, the average value of the apparent binding constant (K(LB): 3.907 X 10(6) L x mol(-1)), and thermodynamics parameters (enthalpy change delta H: -13.43 kJ x mol(-1), entropy change delta S: 81.90 J x K(-1) and standard Gibbs free energy change delta G0: -38.34 kJ x mol(-1)) were calculated, and the amounts of binding sites (n: 1.042)were measured. The fluorescence quenching mechanism of BSA after cefuroxime axetil was added was discussed. BSA was bound with cefuroxime axetil and formed a new compound. The quenching belonged to static fluorescence quenching. The thermodynamic parameters agree with delta H approximately 0, delta S > 0 and delta G0 < 0, and the binding reaction is mainly entropy-driven and electro-static interaction force plays a major role in the reaction. The maximum emission wavelength of Tyr and Trp had an obvious red shift in the synchronous fluorescence spectra, the fluorescence emission wavelength of two peaks had a blue shift in the three-dimensional fluorescence spectrum of BSA in the presence of cefuroxime axetil and the maximum absorbtion wavelenghs of three systems in the UV-Vis absorption spectra were obviously different. These showed that the changes in the micro-environment of Tyr and Trp and demonstrated that the conformation of BSA changed as cefuroxime axetil had been added. This provides important information for discussing the configuration modification of BSA because of the added cefuroxime axetil, and for elucidating the pharmacological effects of cefuroxime axetil and biological effects in the organism.
Wu
Gang-ke
GK
Clinical Laboratory, Central Hospital of Jingzhou, Jingzhou 434020, China. jyk1015@126.com
Yan
Cheng-nong
CN
Liu
Yi
Y
D000042:Absorption; D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis / Q000737:chemistry*; D002417:Cattle; D002444:Cefuroxime / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry; D019277:Entropy; D015394:Molecular Structure; D011485:Protein Binding; D011487:Protein Conformation; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature; D013816:Thermodynamics
D000900:Anti-Bacterial Agents; D012710:Serum Albumin, Bovine; D002444:Cefuroxime; C040738:cefuroxime axetil