Solid-state NMR spectroscopy of microcrystalline proteins

Abstract In this chapter, we discuss recent advances in solid-state magic angle spinning (MAS) NMR spectroscopy of microcrystalline proteins. We review approaches for isotopic labelling and microcrystalline sample preparation and multidimensional MAS NMR correlation experiments for resonance assignment and distance restraint determination as well as the measurement of chemical shift and dipolar tensors. These experiments include heteronuclear 13C/15N-, 1H- and 19F- based techniques in the regimes of moderate (less than 60 kHz) as well as high (greater than 60 kHz) MAS frequencies. We present examples of applications of these techniques to determine 3D structures of proteins and their interactions with water, small-molecule ligands, other proteins as well as nucleic acids. The recent advances in high field magnet and MAS NMR probe technologies open new avenues for methods development and applications to larger proteins and biological assemblies intractable by conventional methods.