High-resolution methyl edited GFT NMR experiments for protein resonance assignments and structure determination

J Biomol NMR. 2010 Nov;48(3):137-45. doi: 10.1007/s10858-010-9444-6. Epub 2010 Sep 14.


Three-dimensional (3D) structure determination of proteins is benefitted by long-range distance constraints comprising the methyl groups, which constitute the hydrophobic core of proteins. However, in methyl groups (of Ala, Ile, Leu, Met, Thr and Val) there is a significant overlap of ¹³C and ¹H chemical shifts. Such overlap can be resolved using the recently proposed (3,2)D HCCH-COSY, a G-matrix Fourier transform (GFT) NMR based experiment, which facilitates editing of methyl groups into distinct spectral regions by combining their ¹³C chemical shifts with that of the neighboring, directly attached, ¹³C nucleus. Using this principle, we present three GFT experiments: (a) (4,3)D NOESY-HCCH, (b) (4,3)D ¹H-TOCSY-HCCH and (c) (4,3)D ¹³C-TOCSY-HCCH. These experiments provide unique 4D spectral information rapidly with high sensitivity and resolution for side-chain resonance assignments and NOE analysis of methyl groups. This is exemplified by (4,3)D NOESY-HCCH data acquired for 17.9 kDa non-deuterated cytosolic human J-protein co-chaperone, which provided crucial long-range distance constraints for its 3D structure determination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Carbon Isotopes
  • Deuterium
  • Escherichia coli / chemistry
  • Escherichia coli / growth & development
  • Fourier Analysis*
  • Humans
  • Isoleucine / chemistry
  • Leucine / chemistry
  • Methylation
  • Molecular Sequence Data
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Protein Structure, Quaternary*
  • Proteins / chemistry*
  • Valine / chemistry


  • Carbon Isotopes
  • Proteins
  • Isoleucine
  • Deuterium
  • Leucine
  • Valine