Resonance assignment and structure investigation by high resolution H-detected solid-state NMR under ultra-fast MAS: from mycrocrystalline proteins to large protein assemblies

We present an overview of our recent advances using ultra-fast (60 kHz) magic-angle spinning (MAS) solid-state NMR spectroscopy and high magnetic fields. These include: (a) the use of deuterated/fully back-exchanged protein samples, which enable the acquisition of high resolution and sensitivity spectra with 1H detection, (b) the design of a suite of scalar-based correlations for resonance assignment of backbone HN, N, C' and C and side-chains C , (c) the α β measurements of site-specific 1H-1H distance restraints using 3D NMR methods. These experiments enable the rapid assignment and the fast determination of the fold of medium-sized proteins, and open the way to the establishment of intermolecular contacts in larger protein assemblies. Examples are presented from microcrystalline and non-crystalline domains and assemblies from E. Coli DNA polymerase, as well as for sedimented viral capsids.