Solid-state NMR spectroscopy of 10% 13C labeled ubiquitin: spectral simplification and stereospecific assignment of isopropyl groups.
We describe the simplification of 13C-13C correlation spectra obtained from a microcrystalline protein sample expressed on a growth medium of 10% fully 13C labeled glucose diluted in 90% natural abundance glucose as compared to a fully labeled sample. Such a labeling scheme facilitates the backbone and side-chain resonance assignment of Phe, Tyr, His, Asp, Asn, Ile, Lys and Pro and yields an unambiguous stereospecific assignment of the valine Cgamma1, Cgamma2 13C resonances and of Leucine Cdelta2.
Schubert
Mario
M
ETH Zurich, Physical Chemistry, 8093, Zurich, Switzerland.
Manolikas
Theofanis
T
Rogowski
Marco
M
Meier
Beat H
BH
D002247:Carbon Isotopes / Q000737:chemistry*; D007553:Isotope Labeling / Q000379:methods*; D007930:Leucine / Q000737:chemistry; D015394:Molecular Structure; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins; D013237:Stereoisomerism; D025801:Ubiquitin / Q000737:chemistry*; D014633:Valine / Q000737:chemistry
D002247:Carbon Isotopes; D011506:Proteins; D025801:Ubiquitin; D007930:Leucine; D014633:Valine