Assignment of the backbone resonances for microcrystalline ubiquitin.
Site-specific assignments for the solid-state NMR spectra of uniformly 13C,15N-enriched ubiquitin are described. The assignments are derived from three three-dimensional 15N-13C-13C correlation spectra collected at 400 MHz on microcrystalline material. A few residues (the loop near Threonine 9 and the C-terminal fragment) were missing and correspond to regions previously reported to be mobile on the basis of X-ray crystallography and solution NMR studies. A few additional sites exhibit shifts that differ from previously reported solution NMR assignments. Nonetheless, these de novo assignments indicate close agreement between the chemical shifts observed in solution and those in microcrystalline or precipitated solids. The methods utilized are likely to be generally applicable for other noncrystalline, nonsoluble proteins.
Igumenova
Tatyana I
TI
Department of Chemistry, Columbia University, New York, 10027 USA.
Wand
A Joshua
AJ
McDermott
Ann E
AE
D002247:Carbon Isotopes; D003460:Crystallization; D018360:Crystallography, X-Ray; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008958:Models, Molecular; D009587:Nitrogen Isotopes; D025801:Ubiquitin / Q000737:chemistry*
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D025801:Ubiquitin