Study on the binding of chloroamphenicol with bovine serum albumin by fluorescence and UV–vis spectroscopy

The binding of chloroamphenicol (CPC) to bovine serum albumin (BSA) at 296 K, 303 K, and 310 K by fluorescence and UV–visible absorption spectroscopy were investigated under imitated physiological conditions. The experimental results showed that the fluorescence quenching mechanism between CPC and BSA was combined quenching (dynamic and static quenching) procedure at low CPC concentration, or a dynamic quenching procedure at high concentrations. The binding constants, binding sites and the corresponding thermodynamic parameters of the interaction system were calculated. According to Förster non-radiation energy transfer theory, the binding distance between CPC and BSA was calculated to be 3.02 nm. Both synchronous fluorescence and FT-IR spectra confirmed the interaction, and indicated the conformational changes of BSA. The effects of some common metal ions Ca2+, Ni2+, Mg2+, Fe2+, and Cu2+ on the binding constant between CPC and BSA were examined. Furthermore, we investigated the possible sub-domains on BSA that bind CPC by displacement experiments.

The interaction between chloroamphenicol (CPC) and bovine serum albumin (BSA) was studied by fluorescence and UV–visible spectroscopy. The quenching mechanism, binding constants and binding distance were obtained. The effects of some common metal ions on the binding constant between CPC and BSA were examined. What’s more, we investigated the possible sub-domains on BSA that bind CPC by displacement experiments.Figure optionsDownload as PowerPoint slideHighlights
► We explored the interaction of BSA and CPC by spectroscopic methods.
► The fluorescence quenching mechanism is combined quenching.
► The binding constants and binding sites were calculated.
► The synchronous fluorescence spectra indicated that the conformation of BSA has been changed.
► The displacement experiment indicated that CPC does bind at the region of site I.