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

Article

Abstract

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.

Keywords

NOESY TOCSY GFT NMR Methyl-edited NOESY Protein structure 

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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