title: Investigation on the pH-dependent binding of benzocaine and lysozyme by fluorescence and absorbance.
abstract: The interaction mechanism between benzocaine (BZC) and lysozyme (Lys) has been investigated by fluorescence, synchronous fluorescence, ultraviolet-vis (UV) absorption spectra, and three-dimensional fluorescence (3-D) in various pH medium. The observations of fluorescence spectra were mainly rationalized in terms of a static quenching process at lower concentration of BZC (C(BZC)/C(Lys)<9) and a combined quenching process at higher concentration of BZC (C(BZC)/C(Lys)>9) at pH 7.4 and 8.4. However, the fluorescence quenching was mainly arisen from static quenching by complex formation in all studied drug concentrations at pH 3.5. The structural characteristics of BZC and Lys were probed, and their binding affinities were determined under different pH conditions (pH 3.5, 7.4, and 8.4). The results indicated that the binding abilities of BZC to Lys decreased at the pH below and above the simulative physiological condition (pH 7.4) due to the alterations of the protein secondary and tertiary structures or the structural change of BZC. The effect of BZC on the conformation of Lys was analyzed using UV, synchronous fluorescence and three-dimensional fluorescence under different pH conditions. These results indicate that the binding of BZC to Lys causes apparent change in the secondary and tertiary structures of Lys. The effect of Zn(2+) on the binding constant of BZC with Lys under various pH conditions (pH 3.5, 7.4, and 8.4) was also studied.
authors.affiliation: College of Chemistry and Chemical Engineering, Luoyang Normal University, Luoyang 471022, China.
authors.forename: Shihui
authors.initials: S
authors.lastname: Li
authors.forename: Daojin
authors.initials: D
authors.lastname: Li
mesh_terms: D000777:Anesthetics / Q000737:chemistry; D000818:Animals; D001566:Benzocaine / Q000737:chemistry*; D002645:Chickens; D015195:Drug Design; D000431:Ethanol / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D021621:Imaging, Three-Dimensional / Q000379:methods; D008968:Molecular Conformation; D009113:Muramidase / Q000737:chemistry*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods
chemical_list: D000777:Anesthetics; D000431:Ethanol; D009113:Muramidase; D001566:Benzocaine