Three-dimensional 13C shift/1H-15N coupling/15N shift solid-state NMR correlation spectroscopy.
Triple-resonance experiments capable of correlating directly bonded and proximate carbon and nitrogen backbone sites of uniformly 13C- and 15N-labeled peptides in stationary oriented samples are described. The pulse sequences integrate cross-polarization from 1H to 13C and from 13C to 15N with flip-flop (phase and frequency switched) Lee-Goldburg irradiation for both 13C homonuclear decoupling and 1H-15N spin exchange at the magic angle. Because heteronuclear decoupling is applied throughout, the three-dimensional pulse sequence yields 13C shift/1H-15N coupling/15N shift correlation spectra with single-line resonances in all three frequency dimensions. Not only do the three-dimensional spectra correlate 13C and 15N resonances, they are well resolved due to the three independent frequency dimensions, and they can provide up to four orientationally dependent frequencies as input for structure determination. These experiments have the potential to make sequential backbone resonance assignments in uniformly 13C- and 15N-labeled proteins.
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Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania, 19104, USA.
Opella
S J
SJ
D002247:Carbon Isotopes; D005998:Glycine / Q000031:analogs & derivatives* / Q000737:chemistry; D006860:Hydrogen Bonding; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010455:Peptides / Q000737:chemistry*; D011522:Protons; D017421:Sequence Analysis; D013113:Spin Labels
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D010455:Peptides; D011522:Protons; D013113:Spin Labels; D005998:Glycine; C006368:aceturic acid