Spectroscopic and molecular modelling study of binding mechanism of bovine serum albumin with phosmet.

Abstract

Phosmet exerts its neurotoxicity by inhibiting acetylcholinesterase that catalyzes the degradation of acetylcholine (a neurotransmitter). Serum proteins are known to influence the biodistribution of various endogenous and exogenous compounds. In the present study, the binding interactions of phosmet with bovine serum albumin (BSA) was investigated to determine the free concentration of phosmet for its neurotoxicity. The binding mechanism was studied using fluorescence, UV-Vis absorption spectroscopy, circular dichroism (CD), and molecular docking techniques. UV-Vis absorption data showed an increase in absorbance of BSA upon binding with phosmet with a slight red-shift in the peak around 280nm. Intrinsic fluorescence of BSA was quenched in the presence of phosmet. The quenching was observed to be inversely correlated to the temperature that indicated the formation of ground state non-fluorescent complex (static quenching). Binding constant values and n values for the binding of phosmet with BSA at three different temperatures confirmed non-covalent binding interactions with a single set of equivalent binding sites. Thermodynamic parameters ∆G (-137.40±3.58kJmol-1); ΔH (-16.33±5.28kJmol-1) and ΔS(-469±12.45kJmol-1) confirmed that the binding was spontaneous and non-covalent interactions like electrostatic, hydrogen bonding and van der Waals forces played an important role in the binding. The CD data indicated the conformational change in BSA upon binding with phosmet which resulted in a change in the melting temperature. Molecular docking presented the binding model for BSA-phosmet complex and displayed that non-covalent interactions played a significant role in the binding mechanism.