Gimenez, Clara: Production of Kindlin-2 F0 Domain for Solid-State NMR Structural Biology

Title: Production of Kindlin-2 F0 Domain for Solid-State NMR Structural Biology

Name: Clara Gimenez

Major: Biomedical Engineering

School affiliation: School of Engineering

Programs: Aresty Summer Science Program

Other contributors: Andrew J. Nieuwkoop and Jacqueline Perodeau

Abstract: Kindlin-2 (K2) is a protein known for mediating cellular chemotaxis by activating integrin. The mechanism of activation involves phosphatidyinositol phosphate (PIP) binding by both the F0 and PH domains (Fig. 1), but the role of each domain is unclear. Characterizing how each domain binds to its PIP ligand and interacts with the bulk membrane is the first step in unraveling the activation cascade. The Nieuwkoop laboratory focuses on applying solid-state nuclear magnetic resonance (ssNMR) techniques to the study of lipid bilayers and the protein that interact with them, including kindling-2.To study PIP binding, the PH and F0 domains were expressed via polymerase chain reaction (PCR) cloning process. Preliminary results found that by refining protocols high protein expression levels were yielded, setting the framework for future ssNMR experiments.