A spectroscopic, molecular docking and molecular dynamic simulation study on the interaction of human hemoglobin with 2,4-dichlorophenoxyacetic acid

Abstract

2,4-Dichlorophenoxyacetic acid (2,4-D) is a systemic herbicide widely used to control dicotyledonous weeds. The general population is routinely exposed to 2,4-D due to consumption of contaminated food and water. 2,4-D is known to damage cellular components in human erythrocytes. This study investigated in detail the interaction of 2,4-D with human hemoglobin (Hb), the major protein in erythrocytes (>95%), and characterized the binding properties utilizing multi-spectrometric and in silico techniques. The UV-visible spectra suggested that 2,4-D interacts with Hb. The fluorescence quenching studies at three different temperatures further showed the binding of 2,4-D to Hb and the formation of a ground-state complex. The results indicated that 2,4-D binds spontaneously to a single moderate-affinity binding site on Hb. Furthermore, the binding process involved van der Waals forces and hydrogen bonding. Circular dichroism and synchronous fluorescence spectra showed that the binding of 2,4-D altered the conformation of Hb and decreased the polarity around its tryptophan residues. 2,4-D binding inhibited the inherent esterase activity of Hb. Computational analysis demonstrated that the Hb-2,4-D complex was stable and identified the amino acid residues at the binding site. Thus, 2,4-D interacts with Hb, modifies the protein conformation and consequently impairs its functions.

Keywords: 2,4-D; Hemoglobin; circular dichroism; esterase activity; fluorescence quenching; in silico.