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Amino Acid Selective Labeling and Unlabeling for Protein Resonance Assignments

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Isotope labeling in Biomolecular NMR

Part of the book series: Advances in Experimental Medicine and Biology ((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.

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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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Authors and Affiliations

  1. NMR Research Centre, Indian Institute of Science, Bangalore, 560012, India

    Garima Jaipuria, B. Krishnarjuna, Somnath Mondal, Abhinav Dubey & Hanudatta S. Atreya

  2. Solid State and Structural Chemistry Unit, Indian Institute of Science, Bangalore, 560012, India

    Garima Jaipuria, Somnath Mondal & Hanudatta S. Atreya

  3. IISc Mathematics Initiative, Indian Institute of Science, Bangalore, 560012, India

    Abhinav Dubey

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  1. Garima Jaipuria
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  2. B. Krishnarjuna
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Correspondence to Hanudatta S. Atreya .

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    Hanudatta S. Atreya

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Jaipuria, G., Krishnarjuna, B., Mondal, S., Dubey, A., Atreya, H.S. (2012). Amino Acid Selective Labeling and Unlabeling for Protein Resonance Assignments. In: Atreya, H. (eds) Isotope labeling in Biomolecular NMR. Advances in Experimental Medicine and Biology, vol 992. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-4954-2_6

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