Expanding our understanding of protein interactions in the cytoplasmic interface of the type four a pili
I want to investigate how the motor proteins responsible for pilus function interact with otherT4aP components. In P. aeruginosa there are two major motor proteins, PilB and PilT, responsible for pilus extension and retraction respectively. I will use co-purification and co-crystallization methods to identify protein-protein interaction surfaces. To this point, I have successfully created a protein expression plasmid library containing various constructs PilB, PilT, PilC, and PilM from various Pseudomonas strains. I also identified protein expression conditions for various of these protein constructs. In addition, I expressed and purify a small domain of PilB and set up screening for crystallographic conditions. Im working on identifying a fast and easy method to identify protein interactions. In addition, I m working in preparing a second plasmid library that will be used for the purpose of identifying novel protein-protein interactions using affinity purification mass spectrometry.