Effect of Hydrogen Peroxide on Hydrolysis of Proteins

Abstract

Hydrogen peroxide (H2O2) is the active ingredient in over‐the‐counter whitening strips for teeth. It has previously been shown to damage certain proteins by making them more susceptible to hydrolysis by proteases. The goal of this project was to characterize the hydrolysis of various proteins with H2O2. Albumin protein was mixed with different concentrations of H2O2 for one hour. The protease trypsin was added and portions were removed and immediately mixed with trichloroacetic acid to precipitate either unhydrolyzed proteins or proteins that were minimally hydrolyzed. After centrifugation, the supernatant, which contains small protein fragments that were released by hydrolysis, was collected and; the total amount was measured using the Lowry assay. The results indicate that even without the protease, there is significant hydrolysis with peroxide and none when the proteins were mixed with water. The amount of hydrolysis correlated with the amount of peroxide. In addition, the amount of hydrolysis in the presence of H2O2 with trypsin greatly exceeded that when only trypsin was present. It was important in this method to inactivate peroxide and efficacy of different metals were tested. This effect was further characterized using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE). These results suggest that hydrogen peroxide; even at the concentrations used in whitening strips, can damage protein by promoting hydrolysis. This work was supported by Research and Professional Development Grant from Stockton University.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.