Investigation on potential enzyme toxicity of clenbuterol to trypsin

Clenbuterol (CLB) is a kind of β2-adrenergic agonists which was illegally used as feed additives nowadays. The toxic interaction of CLB with trypsin, an important digestive enzyme, was studied in vitro using multi-spectroscopic methods and molecular modeling methods. CLB was proved to bind with trypsin in S1 pocket, forming a complex driven by the dominant force of H-bond. The binding constant was calculated to be 1.79887 × 105 L mol−1 at 289 K and 0.32584 × 105 L mol−1 at 310 K, respectively. The skeleton of trypsin became loosened and unfolded with the amino residues microenvironment changed. The secondary and tertiary structure of trypsin also varied. Molecular modeling studies illustrated specific display of the binding information and explained most of the experiment phenomena. The binding site of CLB induced the fluorescence quenching as well as inhibition of enzyme activity of trypsin. The study confirmed that CLB had potential toxicity on both the structure and function of trypsin and the effects enhanced with the increasing concentration of CLB.

Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► Interaction mechanism between clenbuterol and trypsin was investigated. ► Multi-spectroscopic methods and molecular modeling methods were applied. ► The structure and function (enzyme activity) of trypsin were both affected by CLB.