Getting to the “CARL” building where my supervising post-doc works was a pain this morning. I had actually come to the Duke Medical Campus in Downtown Durham earlier in the year, so I was acquainted with the tortuous paths, high-rising brick buildings, and construction sites. Needless to say, I still got lost. Knowing that I’d get lost, I came to the lab one hour early, wandered around for thirty minutes, and (surprisingly) ran into my supervisor, Dr. Masoudi. Promptly, he directed to me to the lab’s manager with whom I signed some confidentiality paperwork. The lab is tucked at the top level of the building, where a plethora of groups are working on biochem projects. My specific lab, the Lefkowitz lab, deals with G-Protein Coupled Receptors that act as intercellular communication devices in eukaryotes (animal cells). These critical receptors come in thousands of different forms but they all work by wrapping around the cell membrane seven times. A “binding site” occupies the end of the receptor outside of the cell while a G-protein – a protein composed of three primary parts that can be ejected to communicate intracellular messages – is connected to the receptor on the inside of the cell. Whenever a unique body binds to the receptor outside of the cell, a “conformational” change occurs where the positioning of the macromolecule slightly alters and releases the G-protein. This complex process is the same process that cells undergo for nearly 40% of our prescribed medicine. The G-protein coupled receptor plays a colossal role in human health, and I can’t wait to get started in the lab!
Today’s labwork began with the expression of beta-2 adrenergic receptors in insect cells. The cells were placed in a solution that expedites receptor formation. The flasks holding these cells were put in a massive centrifuge – a device that spins vessels at high speeds to separate insoluble particles. We poured out the solution, extracted the cells while stabilizing their pH with a buffer solution, and labelled each of our solution-filled flasks. Dr. Masoudi emphasized that everything in the lab must be labeled. We used a smaller centrifuge (going at 4000 rpm!) to separate the insect cells from the buffer solution. Using ethanol and dry ice (the lab ran out of liquid nitrogen), we flash-froze our vessels and later placed our rack of flasks in a massive freezer. The cells in these flasks will eventually be used when the receptors are needed for X-ray crystallography. After our lunch break, Dr. Masoudi and I went over to a lab-wide meeting where different project leaders explicate the results from the past week. Besides X-ray crystallography, other researchers only a couple doors down use cryogenic electron microscopy to better understand the structures of these receptors.
Everyone in the lab is incredibly kind. One researcher called “Bullet” gave up his set of pipets so I could use them later on in the internship. Dr. Masoudi works with another researcher, Li Yin, who was kind enough to give up her work bench and desk so I could use it for labwork. Overall, the lab’s atmosphere is positive, but what I found to be most surprising is that this space is highly diverse; a large majority of the researchers here are first-generation immigrants. Dr. Masoudi just so happens to be a first-generation Iranian immigrant like my mom! It’s been such a phenomenal experience so far, and I’m so grateful for the opportunity to witness some crucial work in the field of biochemistry. Can’t wait for tomorrow!
