Visualizing interactions between RAGE and S100A12 binding and signaling with YFP and CFP

Shaan Shirazi, Verrazzano Class of 2025, completed major in Biology

I was looking for research and was accepted to work in Dr. Rupal Gupta’s lab. There was definitely a learning curve for me, as the projects involved relied heavily on topics in biology and biochemistry that I hadn’t yet learned.

But from the very beginning, I was fascinated by the work being done on proteins and how the experiments conducted could help further our knowledge on the intricate systems that help our bodies function. This was a big motivating factor that kept me coming back and working in the lab. I loved how the work I did would actually help us to work towards a goal of publishing our findings on different proteins and receptors in our bodies, furthering society’s knowledge on this topic.

My capstone covers how my research aims to understand how a protein in our body named S100A12 interacts with its respective receptor called RAGE. Using a technique called fluorescent energy transfer microscopy, it uses a fluorescent protein that we attached to the receptors to visually observe and detect the interactions between S100A12 and RAGE. S100A12 is normally produced by our immune system, and is known to signal immune responses via RAGE, such as inflammation. S100A12 is observed in patients suffering inflammation from diseases such as chronic kidney disease and rheumatoid arthritis.

I found it challenging yet fun learning how to operate different machinery such as the thermocycler to replicate DNA via polymerase chain reaction (PCR) or the gel electrophoresis system to confirm the presence of DNA. I began my research with a basic knowledge on their function and purpose and came out far more experienced and learned. Experiences like that actually benefited me in my biology classes, when these techniques were discussed and tested on.

Working on this project I was very surprised at the time many procedures took, as I quickly learned that science requires a lot of time and patience to get results, and that I always have to brace myself for failure as that is much more common than success. In my capstone paper I detail in my results section the many hiccups I encountered when working towards my goal, and every time I learned to stand back and assess what went wrong and adjust my methods accordingly.

I began this project in my sophomore year, and I have since heavily expanded upon my research on this specific topic involving the S100A12 protein, testing different mutants such as S100A12E31A and S100A12E31AI47A’s ability to bind metal under different conditions. Overall in addition to the scientific knowledge I have gained working on this project, more importantly, I also learned a lot of good skills and habits such as patience, work ethic, and critical thinking.