Discipline: Chemistry and Chemical Sciences
Subcategory: Biochemistry (not Cell and Molecular Biology and Genetics)
Room: Exhibit Hall
Victoria Ogbeifun - Missouri State University
Co-Author(s): Natasha DeVore, Missouri State University, MO
Cytochrome P450s (CYPs) are a family of hemeproteins that are vital to breaking down foreign materials and compounds within mammals. Canine CYP1A2 and canine CYP2E1 are cytochrome P450 enzymes that are specifically found in dogs. The function of these enzymes is to assist with the metabolism of xenobiotics in dogs. Humans have their own versions of both CYP1A2 and CYP2E1. It is these specific enzymes that are accountable for the metabolic differences between dogs and humans when consuming things such as theobromine and caffeine, which are both found in chocolate. Humans seem to tolerate these compounds when they are metabolized, however, dogs tend to experience noxious effects. This poses the question of what are the differences that exist between the canine version of these enzymes and their human counterparts. The goal of this study is to examine the protein activity of canine CYP1A2 and canine CYP2E1 and compare those properties to the equivalent human enzymes. For this project, I will be working on expressing the proteins in E. coli bacteria in order to purify them through nickel affinity chromatography. After purification, the purity will be measured using UV/vis spectroscopy and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Once the proteins are purified, the metabolic activity will be measured through a metabolic assay using high-pressure liquid chromatography (HPLC) to quantify the metabolites produced. These methods will be used to test the hypothesis that there is a significant difference in the metabolic activity of the canine CYP1A2 and CYP2E1 enzymes when compared to the human CYP1A2 and CYP2E1. The results will indicate a purified canine protein and show the metabolic activity that the canine CYP1A2 and CYP2E1 exhibit for caffeine. This study will provide an initial comparison directly at the enzyme level between the breakdown of xenobiotics in canines and humans. Future research involves investigating the metabolism of other compounds known to have substantially different metabolism properties between canines and humans.
Funder Acknowledgement(s): I would like to thank Dr. Obafemi Ajayi-Tayo for her work in the LSAMP program and dedication to helping students succeed in STEM.I would also like to thank Dr. DeVore for all her help, encouragement, and support.
Faculty Advisor: Natasha DeVore, NDeVore@MissouriState.edu
Role: First, I began to read literature and understand precisely the effect that cytochrome P450 enzymes have on humans and Canines. I also examined how caffeine and theobromine are metabolized differently in humans and canines. I also worked on the protein purification by lysing the cells through sonification and centrifugation and preparing them to be run through nickel affinity chromatography. Finally, after the purification, I will be involved in measuring the purity of the proteins and quantifying the metabolites produced.