Exploring Phase Separation of Endocytic Proteins

Students

Faculty

Professor of Chemistry

Project Summary

My PURM experience in the Baumgart lab was an exciting opportunity to gain real experience working in a biochemistry laboratory environment.

Our project focused on physical interactions between a few different proteins that are involved in endocytosis, the process by which cells bend their membrane inwards to envelop outside material. We know that interactions between proteins that contain SH3 domains and proteins that contain PRM sections lead to a phenomenon known as phase separation, in which the proteins cluster and form a droplet separate from the larger solution, like an oil droplet in water. The protein endophilin, which has an SH3 domain, is important to change the membrane’s shape during endocytosis, and the protein lamellipodin has a section with multiple PRM’s and is known to interact with endophilin during endocytosis.

I spent the summer helping to purify these proteins from bacteria and investigate their interactions. Firstly, I learned several techniques important for biochemistry research, including operating our lab’s fast protein liquid chromatography (FPLC) machine, which purifies the proteins we want from other proteins present in the bacteria. A significant portion of the summer was spent perfecting our FPLC procedure, because the proteins we attempted to purify were particularly difficult to isolate. I also learned some techniques specific to membrane biochemistry, such as preparing GUV’s, membrane spheres that mimic actual cell membranes, as well as some data-collecting techniques such as DLS which uses light scattering to ascertain the size of particles in a solution.

Besides the actual lab experience, interacting with the other group members provided a view into the world of research and helped me understand possible directions for my future in research. I came out of my research project with practical wet lab experience, a better understanding of topics covered in my lecture and lab courses, and a strengthened interest in pursuing research in the future.