I started working on this project summer of 2014 through ASSIP (Aspiring Scientists Summer Internship Program); when I was filling out the application they had the list of mentors and the description of their project. At that point my mom was going through chemotherapy for breast cancer, therefore, when I read the description of Dr. Paige’s project I was really interested and I chose him as a first mentor. I was chosen and started working on this project since then. This project is personally related to me as well as it is related to my major and future goals. I am a pre-med student at GMU, therefore, anything related to health and human body would interest me, catch my attention, and I would do anything in my ability to learn more about the field.
The hypothesis of this experiment is: Whether the persistent inflammation associated with emphysema due to the inactivation of the LTA4H enzyme? To test this hypothesis, we used L-alanine p-nitroanilide (Ala-pNA) as a substrate to mimic PGP and determine the rate of aminopeptidase activity. Release of the pNA reporter group is mediated by the aminopeptidase activity of the LTA4H enzyme. We independently varied the concentration of the substrate in the presence of a constant concentration of the enzyme, 4MDM, and buffer. On the weekly basis I measure the progress of the reaction, each component, enzyme, substrate and drug, are placed into a Biotek UV-Vis plate reader to measure absorption at [lamda]= 405 nm and determine the rate of p-nitroaniline formation, which corresponds to the aminopeptidase activity of the enzyme. Using the data from the UV-Vis plate reader, we calculate the initial velocity (Vo) of the reaction at multiple concentrations of substrate or drug. During the past couple weeks I am trying to get a data that is reusable and constant and based on the last run it looked very constant. The next step is to use different molecule (4MDM).