Journal of Biomolecular NMR

, Volume 52, Issue 2, pp 115–126

Efficient sequential assignments in proteins with reduced dimensionality 3D HN(CA)NH


  • Kousik Chandra
    • NMR Research CentreIndian Institute of Science
  • Garima Jaipuria
    • NMR Research CentreIndian Institute of Science
    • Solid State and Structural Chemistry UnitIndian Institute of Science
  • Divya Shet
    • NMR Research CentreIndian Institute of Science
    • NMR Research CentreIndian Institute of Science

DOI: 10.1007/s10858-011-9598-x

Cite this article as:
Chandra, K., Jaipuria, G., Shet, D. et al. J Biomol NMR (2012) 52: 115. doi:10.1007/s10858-011-9598-x


We present reduced dimensionality (RD) 3D HN(CA)NH for efficient sequential assignment in proteins. The experiment correlates the 15N and 1H chemical shift of a residue (‘i’) with those of its immediate N-terminal (i − 1) and C-terminal (i + 1) neighbors and provides four-dimensional chemical shift correlations rapidly with high resolution. An assignment strategy is presented which combines the correlations observed in this experiment with amino acid type information obtained from 3D CBCA(CO)NH. By classifying the 20 amino acid types into seven distinct categories based on 13Cβ chemical shifts, it is observed that a stretch of five sequentially connected residues is sufficient to map uniquely on to the polypeptide for sequence specific resonance assignments. This method is exemplified by application to three different systems: maltose binding protein (42 kDa), intrinsically disordered domain of insulin-like growth factor binding protein-2 and Ubiquitin. Fast data acquisition is demonstrated using longitudinal 1H relaxation optimization. Overall, 3D HN(CA)NH is a powerful tool for high throughput resonance assignment, in particular for unfolded or intrinsically disordered polypeptides.


Sequence specific resonance assignmentReduced dimensionality NMRProtein structureGFT NMR

Supplementary material

10858_2011_9598_MOESM1_ESM.pdf (332 kb)
Supplementary material 1 (PDF 331 kb)

Copyright information

© Springer Science+Business Media B.V. 2011