Strategies for RNA Resonance Assignment by 13C/15N- and 1H-Detected Solid-State NMR Spectroscopy

Magic angle spinning (MAS) solid-state NMR (ssNMR) is an established tool that can be applied to non-soluble or non-crystalline biomolecules of any size or complexity. The ssNMR method advances rapidly due to technical improvements and the development of advanced isotope labeling schemes. While ssNMR has shown significant progress in structural studies of proteins, the number of RNA studies remains limited due to ssNMR methodology that is still underdeveloped. Resonance assignment is the most critical and limiting step in the structure determination protocol that defines the feasibility of NMR studies. In this review, we summarize the recent progress in RNA resonance assignment methods and approaches for secondary structure determination by ssNMR. We critically discuss advantages and limitations of conventional 13C- and 15N-detected experiments and novel 1H-detected methods, identify optimal regimes for RNA studies by ssNMR, and provide our view on future ssNMR studies of RNA in large RNP complexes.

Download Full-text

Filamentous Bacteriophage Viruses: Preparation, Magic-Angle Spinning Solid-State NMR Experiments, and Structure Determination

Methods in Molecular Biology - Protein NMR ◽

10.1007/978-1-4939-7386-6_4 ◽

2017 ◽

pp. 67-97 ◽

Cited By ~ 4

Author(s):

Omry Morag ◽

Nikolaos G. Sgourakis ◽

Gili Abramov ◽

Amir Goldbourt

Keyword(s):

Solid State ◽

Structure Determination ◽

Solid State Nmr ◽

Magic Angle Spinning ◽

Magic Angle ◽

Filamentous Bacteriophage ◽

Angle Spinning

Download Full-text

Dihedral Angle Measurements for Structure Determination by Biomolecular Solid-State NMR Spectroscopy

Frontiers in Molecular Biosciences ◽

10.3389/fmolb.2021.791090 ◽

2021 ◽

Vol 8 ◽

Author(s):

Patrick C. A. van der Wel

Keyword(s):

Solid State ◽

Structure Determination ◽

Solid State Nmr ◽

Structural Information ◽

Magic Angle Spinning ◽

Magic Angle ◽

Distance Measurements ◽

Challenges And Opportunities ◽

Molecule Interactions ◽

Angle Spinning

In structural studies of immobilized, aggregated and self-assembled biomolecules, solid-state NMR (ssNMR) spectroscopy can provide valuable high-resolution structural information. Among the structural restraints provided by magic angle spinning (MAS) ssNMR the canonical focus is on inter-atomic distance measurements. In the current review, we examine the utility of ssNMR measurements of angular constraints, as a complement to distance-based structure determination. The focus is on direct measurements of angular restraints via the judicious recoupling of multiple anisotropic ssNMR parameters, such as dipolar couplings and chemical shift anisotropies. Recent applications are highlighted, with a focus on studies of nanocrystalline polypeptides, aggregated peptides and proteins, receptor-substrate interactions, and small molecule interactions with amyloid protein fibrils. The review also examines considerations of when and where ssNMR torsion angle experiments are (most) effective, and discusses challenges and opportunities for future applications.

Download Full-text