High-resolution solid-state NMR studies on uniformly [13C,15N]-labeled ubiquitin.
Understanding of the effects of intermolecular interactions, molecular dynamics, and sample preparation on high-resolution magic-angle spinning NMR data is currently limited. Using the example of a uniformly [13C,15N]-labeled sample of ubiquitin, we discuss solid-state NMR methods tailored to the construction of 3D molecular structure and study the influence of solid-phase protein preparation on solid-state NMR spectra. A comparative analysis of 13C', 13Calpha, and 13Cbeta resonance frequencies suggests that 13C chemical-shift variations are most likely to occur in protein regions that exhibit an enhanced degree of molecular mobility. Our results can be refined by additional solid-state NMR techniques and serve as a reference for ongoing efforts to characterize the structure and dynamics of (membrane) proteins, protein complexes, and other biomolecules by high-resolution solid-state NMR.
Seidel
Karsten
K
Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
Etzkorn
Manuel
M
Heise
Henrike
H
Becker
Stefan
S
Baldus
Marc
M
D002247:Carbon Isotopes; D018360:Crystallography, X-Ray; D006639:Histidine / Q000737:chemistry; D008958:Models, Molecular; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D025801:Ubiquitin / Q000737:chemistry* / Q000378:metabolism
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D025801:Ubiquitin; D006639:Histidine