Probing molecular interfaces using 2D magic-angle-spinning NMR on protein mixtures with different uniform labeling.
A general NMR strategy to directly study molecular interfaces under magic angle spinning is introduced. The approach is based on the spectroscopic analysis of uniformly, but heterogeneously, labeled molecular mixtures containing the spin species X and Y (X:Y). For the case of an ((15)N:(13)C) labeled sample, the use of NC, NHC, and NHHC transfers is demonstrated. Applied to the ((13)C:(15)N) labeled dimeric form of the 85 amino acid protein Crh, the NHHC approach reveals a variety of monomer-monomer interactions in the microcrystalline state.
Etzkorn
Manuel
M
Max-Planck-Institute for Biophysical Chemistry, Department of NMR-based Structural Biology, Am Fassberg 11, 37077 Göttingen, Germany.
Böckmann
Anja
A
Lange
Adam
A
Baldus
Marc
M
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D002247:Carbon Isotopes; D019281:Dimerization; D005947:Glucose / Q000737:chemistry; D005998:Glycine / Q000737:chemistry; D007553:Isotope Labeling; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010750:Phosphoproteins / Q000737:chemistry*; D017434:Protein Structure, Tertiary
D001426:Bacterial Proteins; D002247:Carbon Isotopes; C118042:Crh protein, Bacillus subtilis; D009587:Nitrogen Isotopes; D010750:Phosphoproteins; D005947:Glucose; D005998:Glycine