The inside of the cell is a crowded and complex environment that is impossible to duplicate by studying proteins and other molecules in dilute solution. The effects of macromolecular crowding are potentially important to all cellular functions, but until recently studies have been conducted mostly in dilute solution. In-cell Nuclear Magnetic Resonance (NMR) spectroscopy is becoming an important tool in studying proteins under physiologically relevant conditions. In some instances, however, protein signals from leaked protein are seen in the supernatant surrounding the in-cell slurry. I examined how expression levels contribute to protein leakage. I also describe a device and system that provides a controlled environment for NMR experiments in living cells. I have utilized this device to study the expression of the natively disordered protein - synuclein, inside Escherichia coli. In the future, we hope to make progress in using this device to study proteins in eukaryotic cells with NMR
