New Insights into the Toxicity ofn-Butanol to Trypsin: Spectroscopic and Molecular Docking Descriptions

Abstract

n-Butanol has been widely used and its residue exists extensively in the environment. It could lead to conformational and functional changes of trypsin by forming a complex with it. Docking method and spectrographic technique were employed to the study of the complex of trypsin and n-butanol. The fluorescence results indicated that n-butanol can form a complex with trypsin and change the distance between tryptophan and fluorescence quenchers. The conformational changes of trypsin were proved by UV-visible absorption and synchronous fluorescence spectroscopy indicating that n-butanol had little effect on the conformation of trypsin at a low concentration while denatured and coagulated the trypsin at a high concentration. The binding site was displayed by molecular modeling, which gave information about distances and binding forces between n-butanol and trypsin. The results were in accordance with spectroscopic experiments. Besides, enzyme activity assay gave the dose-response relationship of n-butanol with trypsin.

Enzyme Inhibition-Based Assay to Estimate the Contribution of Formulants to the Effect of Commercial Pesticide Formulations

“…The general purpose of the methods used to establish mechanisms of enzyme inhibition by toxic substances is to solve two types of problems: to reveal formation of the bond between the enzyme and the inhibitor (formation of a stable complex) and to detect possible conformational changes in the structure of proteins in the presence of toxic substances. For these purposes, various optical techniques, including absorption, fluorescence, and circular dichroism spectroscopy, as well as molecular modeling methods are widely used [47][48][49].…”