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Journal of Biomolecular NMR

, Volume 42, Issue 2, pp 149–154 | Cite as

Chemical shift based editing of CH3 groups in fractionally 13C-labelled proteins using GFT (3, 2)D CT-HCCH-COSY: stereospecific assignments of CH3 groups of Val and Leu residues

  • Ravi Pratap Barnwal
  • Hanudatta S. AtreyaEmail author
  • Kandala V. R. CharyEmail author
Communication

Abstract

We propose a (3, 2)D CT-HCCH-COSY experiment to rapidly collect the data and provide significant dispersion in the spectral region containing 13C–1H cross peaks of CH3 groups belonging to Ala, Ile, Leu, Met, Thr and Val residues. This enables one to carry out chemical shift based editing and grouping of all the 13C–1H cross peaks of CH3 groups belonging to Ala, Ile, Leu, Met, Thr and Val residues in fractionally (10%) 13C-labelled proteins, which in turn aids in the sequence-specific resonance assignments in general and side-chain resonance assignments in particular, in any given protein. Further, we demonstrate the utility of this experiment for stereospecific assignments of the pro-R and pro-S methyl groups belonging to the Leu and Val residues in fractionally (10%) 13C-labelled proteins. The proposed experiment opens up a wide range of applications in resonance assignment strategies and structure determination of proteins.

Keywords

Isotope labeling NMR Automated assignments Sequence specific resonance assignments Stereo-specific resonance assignments 

Notes

Acknowledgements

The facilities provided by National Facility for High Field NMR at TIFR supported by Department of Science and Technology (DST), Department of Biotechnology (DBT), Council of Scientific and Industrial Research (CSIR), and Tata Institute of Fundamental Research, Mumbai, India, and NMR Research Centre at IISc supported by DST and are gratefully acknowledged. HSA acknowledges support from Department of Atomic Energy (DAE) BRNS and DST-SERC research awards. We thank Dr. Yogendra Sharma (CCMB, Hyderabad) for providing us the M-crystallin expression plasmid.

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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  1. 1.Department of Chemical SciencesTata Institute of Fundamental ResearchColaba, MumbaiIndia
  2. 2.NMR Research CentreIndian Institute of ScienceMalleshwaram, BangaloreIndia

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