A Targeted DNP Approach to Study the Activity of Membrane Proteins in Whole Bacterial Cells

While solid-state NMR spectroscopy is the method of choice when it comes to studying the
dynamics of proteins in heterogeneous complex environments, inherently dilute samples like membrane proteins demand a more sensitive technique. In this study, we use a targeted DNP approach to study membrane proteins in whole cells where an unnatural amino acid (UAA) is installed into the region of interest on the protein. In the context of whole bacterial cells, we install a UAA into the protein Ail and target it with a DNP polarization agent using bio-orthogonal chemistry. This method specifically enhances the NMR signals of the region of interest over the cellular background enabling us to study protein-lipid interactions in the native context..

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