Unambiguous correlations of backbone amide and aliphatic gamma resonances in deuterated proteins.
Two 3D NMR pulse sequences that correlate aliphatic gamma carbon resonance frequencies to amide proton and nitrogen chemical shifts in perdeuterated proteins are presented. The HN(COCACB)CG provides only interresidue connectivities (NH(i) and Cgamma(i-1)) while the HN(CACB)CG detects both the inter- and intraresidue (NH(i) and Cgamma(i) or Cgamma(i-1)) correlations. These two experiments are useful for sequential assignments and the identification of residue type from the Cgamma shifts. Spectra acquired on a perdeuterated 53-kDa protein illustrate the sensitivity and utility of these experiments.
McCallum
S A
SA
Department of Biochemistry, University of Virginia School of Medicine, Charlottesville, Virginia 22908, USA.
Hitchens
T K
TK
Rule
G S
GS
D000577:Amides / Q000032:analysis*; D003903:Deuterium / Q000032:analysis*; D005982:Glutathione Transferase / Q000032:analysis; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation*; D009682:Magnetic Resonance Spectroscopy / Q000295:instrumentation*; D011506:Proteins / Q000032:analysis*
D000577:Amides; D011506:Proteins; D003903:Deuterium; D005982:Glutathione Transferase