Long-range 19F-15N distance measurements in highly-13C, 15N-enriched solid proteins with 19F-dephased REDOR shift (FRESH) spectroscopy

Daniel T. Graesser, Benjamin J. Wylie, Andrew J. Nieuwkoop, W. Trent Franks, Chad M. Rienstra

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We present a novel rotational-echo double resonance (REDOR) method for detection of multiple 19F-15N distances in solid proteins. The method is applicable to protein samples containing a single 19F label, in addition to high levels of 13C and 15N enrichment. REDOR dephasing pulses are applied on the 19F channel during an indirect constant time chemical shift evolution period on 15N, and polarization is then transferred to 13C for detection, with high-power 1H decoupling throughout the sequence. This four-channel experiment reports site-specifically on 19F- 15N distances, with highly accurate determinations of ∼5 Å distances and detection of correlations arising from internuclear distances of at least 8 A. We demonstrate the method on the well-characterized 56-residue model protein GB1, where the sole tryptophan residue (Trp-43) has been labeled with 5-19F-Trp, in a bacterial growth medium also including 13C-glucose and 15N ammonium chloride. In GB1, 11 distances are determined, all agreeing within 20% of the X-ray structure distances. We envision the experiment will be utilized to measure quantitative long-range distances for protein structure determination.

Original languageEnglish
Pages (from-to)S129-S134
JournalMagnetic Resonance in Chemistry
Issue numberSUPPL.
StatePublished - Dec 2007


  • C
  • Dipolar recoupling
  • F
  • H
  • Isotopic labeling
  • Magic angle spinning
  • N
  • NMR
  • Protein structure


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