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Amino Acid Selective Labeling and Unlabeling for Protein Resonance Assignments

  • Garima Jaipuria
  • B. Krishnarjuna
  • Somnath Mondal
  • Abhinav Dubey
  • Hanudatta S. AtreyaEmail author
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 992)

Abstract

Structural characterization of proteins by NMR spectroscopy begins with the process of sequence specific resonance assignments in which the 1H, 13C and 15N chemical shifts of all backbone and side-chain nuclei in the polypeptide are assigned. This process requires different isotope labeled forms of the protein together with specific experiments for establishing the sequential connectivity between the neighboring amino acid residues. In the case of spectral overlap, it is useful to identify spin systems corresponding to the different amino acid types selectively. With isotope labeling this can be achieved in two ways: (i) amino acid selective labeling or (ii) amino acid selective ‘unlabeling’. This chapter describes both these methods with more emphasis on selective unlabeling describing the various practical aspects. The recent developments involving combinatorial selective labeling and unlabeling are also discussed.

Keywords

HSQC Spectrum Amino Acid Type Label Amino Acid Selective Label Stereospecific Assignment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

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 and DAE-BRNS research awards. GJ acknowledges fellowship from Council of Scientific and Industrial Research (CSIR). We thank Dr. John Cort, Pacific Northwest National Laboratory, for providing the Ubiquitin plasmid.

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Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  • Garima Jaipuria
    • 1
    • 2
  • B. Krishnarjuna
    • 1
  • Somnath Mondal
    • 1
    • 2
  • Abhinav Dubey
    • 1
    • 3
  • Hanudatta S. Atreya
    • 1
    • 2
    Email author
  1. 1.NMR Research CentreIndian Institute of ScienceBangaloreIndia
  2. 2.Solid State and Structural Chemistry UnitIndian Institute of ScienceBangaloreIndia
  3. 3.IISc Mathematics InitiativeIndian Institute of ScienceBangaloreIndia

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