Cellular Solid-State NMR Applied to Bacterial and Human cells

Abstract

Solid-state NMR (ssNMR) is a method that can be applied to gain atomic-level insight into heterogeneous molecular systems. Over the last 15 years, ssNMR has seen strong methodological and instrumental developments that have allowed for the characterization of complex molecules including membrane proteins, amyloid fibrils or protein biopolymers with remarkable structural accuracy and comprehensiveness. More recently, ssNMR has profited from revolutionary enhancements in sensitivity, mainly due to the advent of Dynamic Nuclear Polarization (DNP) and it has seen significant progress in the field of ssNMR 1H detection. In our contribution we show how to make combined use of such methods to probe biomolecules in bacterial and human cells. Applications include protein translocation and insertion machines in bacteria and extend to membrane associated as well as soluble protein complexes in human cells. We also describe how to combine such ssNMR-based studies with modalities such as cryo-electron tomography (CET) and Fluorescence microscopy to obtain structural insight into cell organization from the atomic to sub-micrometer scale.