Using fluorescence and circular dichroism (CD) spectroscopy to investigate the interaction between di-n-butyl phthalate and bovine serum albumin

Abstract

The interaction between di-n-butyl phthalate (DBP) and bovine serum albumin (BSA) in physiological Tris-HCl buffer at pH 7.4 was investigated by fluorescence quenching technique. By analyzing the fluorescence spectrum and intensity, it was observed that the DBP had a strong ability to quench the intrinsic fluorescence of BSA through a static quenching procedure. The binding constants K and the number of binding sites n of DBP with BSA were calculated to be 0.11 × 10² L·mol⁻¹ and 0.52 at 298 K, respectively. The thermodynamic parameters of enthalpy change (ΔH) and entropy change (ΔS) were also calculated to be positive showing that hydrophobic forces might play a major role in the binding of DBP to BSA. The binding process was spontaneous in which Gibbs free energy change (ΔG) was negative. The distance (r) between the donor (BSA) and acceptor (DBP) was calculated to be 2.02 nm based on Forster’s non-radiative energy transfer theory, which indicated that the energy transfer from BSA to DBP occurs with a high possibility. The synchronous fluorescence, three-dimensional fluorescence, and circular dichroism (CD) spectra showed that the binding of di-n-butyl phthalate to BSA induced conformational changes in BSA. The interaction between DBP and BSA can help researchers better understand the nature of poisons and serve people in the right way with first aid and detoxification.

Keywords:

• Bovine serum albumin (BSA)
• di-n-butyl phthalate (DBP)
• fluorescence spectra
• binding process
• toxicity

Data availability statement

The authors confirm that the original data supporting the findings of this study are available within the article or its supplementary materials.

Additional information

Funding

This work was supported by Shenyang Science and Technology Program (No. 22-322-3-14), National Natural Science Foundation of China (No. 52170163) and Scientific Research Project of Liaoning Provincial Department of Education (No. LJKZ0092).