During this past summer I was given the amazing opportunity to participate in the WAVE fellows program and to work in the Rees lab at the California Institute of Technology in Pasadena. With the Rees group I worked on a project consisting of biochemical and structural analysis of a high affinity methionine ABC transporter in E. coli. Prior to my arrival, the lab had ordered nanobodies isolated from llamas that bound specifically to the MetNI transporter. My goal for the summer was to characterize the binding of the nanobodies to MetNI by assessing conformational dynamics, enzymatic activity, and the binding affinity. I used lanthanide resonance energy transfer (LRET) to determine how the nanobodies were changing the conformation of the transporter by measuring the distances between the two nucleotide binding domains. To supplement the LRET data, I also determined how the nanobodies affected the ATPase activity of the transporter, which is coupled to methionine transport. I also utilized surface plasmon resonance (SPR) experiments to determine how tightly the nanobodies were binding to MetNI. Although I spent countless hours attempting to crystalize the transporter, I was not able to get crystals with good enough diffraction for solving the structure. Despite the lack of structures, I was able to characterize nanobody binding to MetNI using biochemical techniques. I learned a lot of things during the summer including working with membrane proteins and x-ray crystallography.
It was a fun summer escaping the arid climate of Arizona. Pasadena is a cool city with a lot to do within the city, as well as the surrounding areas. It’s only a 45-minute drive from LA, an hour from Santa Monica, and 30 minutes from the San Gabriel Mountains. I spent a majority of my days on the beach, when I wasn’t in the lab. The program offered many social events such as a trip to a Dodgers game, a trip to Santa Monica, and others. There were plenty of workshops geared towards preparing us for graduate school. Overall it was a great summer; I got to do some exciting research, and I got to relax on the beach.