UC San Diego

Solid-state nuclear magnetic resonance (NMR) spectroscopy is a powerful method for the study of membrane proteins under native environments. G protein-coupled receptors (GPCRs) are highly challenging integral membrane proteins that are often hard to study using the available biophysical techniques. Limited information about their structures and dynamics has been elucidated with x-ray crystallography as well as various biochemical techniques, such as Föster resonance energy transfer (FRET), and mutation studies. Obtaining site resolved NMR spectra for GPCRs can help to better understand their structures and behaviors. Here CXC-chemokine receptor 2 (CXCR2) is studied with various NMR tools, and the functional relevance of the obtained protein is verified with ligand binding assays as well as NMR. A comparison of CXCR2 interaction with two of its ligands: interleukin 8 and CXCL5 is also presented. At the same time, methods such as protein perdeuteration, and 1H detection were developed for the study of complex membrane proteins using the oriented-sample solid-state NMR technique. NMR developments are also made on the MerFTC chimeric protein, to help experiments for the more complex CXCR2. Combining various NMR techniques, optimized NMR spectra are presented, providing insights into the structure and function of the CXCR2 protein.