Journal of Biomolecular NMR

, Volume 48, Issue 3, pp 137–145 | Cite as

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

  • Garima Jaipuria
  • Anushikha Thakur
  • Patrick D’SilvaEmail author
  • Hanudatta S. AtreyaEmail author


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.


NOESY TOCSY GFT NMR Methyl-edited NOESY Protein structure 



The facilities provided by NMR Research Centre at IISc supported by Department of Science and Technology (DST), India is gratefully acknowledged. HSA acknowledges support from DST-SERC research award. GJ and AT acknowledge fellowship from Council of Scientific and Industrial Research (CSIR) and UGC, India. PDS acknowledges support from the Wellcome Trust International Senior Research Fellowship in Biomedical Science, WT081643MA. We thank Dr. John Cort, Pacific Northwest National Laboratory, for providing the Ubiquitin plasmid.

Supplementary material

10858_2010_9444_MOESM1_ESM.doc (246 kb)
Supplementary material 1 (DOC 246 kb)


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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.NMR Research CentreIndian Institute of ScienceBangaloreIndia
  2. 2.Solid State and Structural Chemistry UnitIndian Institute of ScienceBangaloreIndia
  3. 3.Chemical Biology ProgrammeIndian Institute of ScienceBangaloreIndia
  4. 4.Department of BiochemistryIndian Institute of ScienceBangaloreIndia

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