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).
