Bacterial Effector Protein: From Cloning to Sample Preparation for Biophysical Studies

Louis Herrera, McKenna Parker, Sofia Gonzalez, Dimitra Keramisanou, Ioannis Gelis

University of South Florida, Department of Chemistry

Pathogenic bacteria use a variety of mechanisms to invade its host, including the use of effector proteins that interact with the host’s cellular pathways and inhibit its immune defense. Understanding bacterial mechanisms for infection give opportunity for developing new antibiotics. Here, we investigate a bacterial effector protein that leads to the inhibition of the NF-kB pathway and shuts down a cell’s inflammatory responses. It belongs to a conserved family of effector proteins, some of which have been previously studied. Common to all members of this family is that they contain a minimal protein fold compared to their corresponding non-effector homologues that is relatively unstable until it forms complexes with host proteins after translocation. We are building experimental methods to effectively clone, express and purify members of this family aiming in preparing samples that can be used for analyzing their chemical and physical properties as well as comparing it to other bacterial effector proteins.