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Solid‐State NMR H–N–(C)–H and H–N–C–C 3D/4D Correlation Experiments for Resonance Assignment of Large Proteins
Author(s) -
Fraga Hugo,
Arnaud CharlesAdrien,
Gauto Diego F.,
Audin Maxime,
Kurauskas Vilius,
Macek Pavel,
Krichel Carsten,
Guan JiaYing,
Boisbouvier Jerome,
Sprangers Remco,
Breyton Cécile,
Schanda Paul
Publication year - 2017
Publication title -
chemphyschem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.016
H-Index - 140
eISSN - 1439-7641
pISSN - 1439-4235
DOI - 10.1002/cphc.201700572
Subject(s) - heteronuclear molecule , chemistry , nuclear magnetic resonance spectroscopy , solid state nuclear magnetic resonance , magic angle spinning , amide , crystallography , protonation , spectroscopy , resonance (particle physics) , molecule , two dimensional nuclear magnetic resonance spectroscopy , nmr spectra database , spectral line , nuclear magnetic resonance , stereochemistry , atomic physics , physics , ion , organic chemistry , quantum mechanics , astronomy
Solid‐state NMR spectroscopy can provide insight into protein structure and dynamics at the atomic level without inherent protein size limitations. However, a major hurdle to studying large proteins by solid‐state NMR spectroscopy is related to spectral complexity and resonance overlap, which increase with molecular weight and severely hamper the assignment process. Here the use of two sets of experiments is shown to expand the tool kit of 1 H‐detected assignment approaches, which correlate a given amide pair either to the two adjacent CO–CA pairs (4D hCOCANH/hCOCAcoNH), or to the amide 1 H of the neighboring residue (3D HcocaNH/HcacoNH, which can be extended to 5D). The experiments are based on efficient coherence transfers between backbone atoms using INEPT transfers between carbons and cross‐polarization for heteronuclear transfers. The utility of these experiments is exemplified with application to assemblies of deuterated, fully amide‐protonated proteins from approximately 20 to 60 kDa monomer, at magic‐angle spinning (MAS) frequencies from approximately 40 to 55 kHz. These experiments will also be applicable to protonated proteins at higher MAS frequencies. The resonance assignment of a domain within the 50.4 kDa bacteriophage T5 tube protein pb6 is reported, and this is compared to NMR assignments of the isolated domain in solution. This comparison reveals contacts of this domain to the core of the polymeric tail tube assembly.