Advances in instrumentation and methodology for solid-state NMR of biological assemblies

Abstract

Many advances in instrumentation and methodology have furthered the use of solid-state NMR as a technique for determining the structures and studying the dynamics of molecules involved in complex biological assemblies. Solid-state NMR does not require large crystals, has no inherent size limit, and with appropriate isotopic labeling schemes, supports solving one component of a complex assembly at a time. It is complementary to cryo-EM, in that it provides local, atomic-level detail that can be modeled into larger-scale structures. This review focuses on the development of high-field MAS instrumentation and methodology; including probe design, benchmarking strategies, labeling schemes, and experiments that enable the use of quadrupolar nuclei in biomolecular NMR. Current challenges facing solid-state NMR of biological assemblies and new directions in this dynamic research area are also discussed.

Chemical Properties Determine Solubility and Stability in βγ‐Crystallins of the Eye Lens

“…For large complexes or insoluble aggregates, such as the amorphous or amyloid aggregates formed in cataract, solidstate NMR spectroscopy with magic angle spinning (MAS) is a useful structural method. [67][68][69][70] This technique can be used either on its own or synergistically with other methods including small-angle X-ray scattering (SAXS), or for larger complexes, electron microscopy. Solid-state NMR has been used to show native-like structure in aggregates of the P23T variant of HγDcrystallin, [55] and to solve the structures of αB-crystallin complexes.…”

“…For large complexes or insoluble aggregates, such as the amorphous or amyloid aggregates formed in cataract, solid‐state NMR spectroscopy with magic angle spinning (MAS) is a useful structural method [67–70] . This technique can be used either on its own or synergistically with other methods including small‐angle X‐ray scattering (SAXS), or for larger complexes, electron microscopy.…”

Structures of Pathological and Functional Amyloids and Prions, a Solid-State NMR Perspective

“…Figure 5A presents various rotor sizes used for biomolecular studies, ranging from 4 mm diameter rotor (10–100 mg sample quantity, routinely used at 11 kHz MAS) to 0.7 mm diameter rotor (0.5–1 mg sample quantity, routinely used at 100 kHz MAS, as illustrated in Figure 5B for HET-s amyloid fibrils). Nowadays, many semi-automatic setups are available as state-of-the-art for commercial SSNMR probes, which are well adapted for the needs of structural biology research ( Martin et al, 2019 ). These technological developments, combined with the increased number of available high-field NMR magnets and advanced protein labeling techniques have considerably improved the analytical capability of SSNMR ( Lacabanne et al, 2019 ).…”

Recent advances in probing host–guest interactions with solid state nuclear magnetic resonance

“…The NMR sensitivity is largely dependent on the Boltzmann energy difference between the magnetic states of the nuclear spin which is determined by the strength of the external magnetic field and the value of the gyromagnetic ratio of the nucleus. High field NMR [20][21][22] is therefore often used to overcome this challenge by increasing this energy gap and also often results in higher resolution by increasing chemical shift dispersion. Dynamic nuclear polarisation (DNP) [23][24][25] and cross polarisation (CP) 26 emerged as powerful approaches to significantly increase sensitivity by polarisation transfer of the high magnetisation of electron spins to nucleus spins, and of higher polarised nucleus spins (e.g.…”