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In-Cell NMR of Intrinsically Disordered Proteins in Prokaryotic Cells

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Intrinsically Disordered Protein Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 895))

Abstract

In-cell NMR, i.e., the acquisition of heteronuclear multidimensional NMR of biomacromolecules inside living cells, is, to our knowledge, the only method for investigating the three-dimensional structure and dynamics of proteins at atomic detail in the intracellular environment. Since the inception of the method, intrinsically disordered proteins have been regarded as particular targets for in-cell NMR, due to their expected sensitivity to the molecular crowding in the intracellular environment. While both prokaryotic and eukaryotic cells can be used as host cells for in-cell NMR, prokaryotic in-cell NMR, particularly employing commonly used protein overexpression systems in Escherichia coli cells, is the most accessible approach. In this chapter we describe general procedures for obtaining in-cell NMR spectra in E. coli cells.

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Acknowledgments

This work was supported in part by the CREST program of the Japan Science and Technology Agency (JST), the Molecular Ensemble Program of RIKEN, Grants-in-Aid for Scientific Research of Priority Areas on “Molecular Soft Interactions Regulating Membrane Interface of Biological Systems” and “Molecular Science for Supra Functional Systems―Development of Advanced Methods for Exploring Elementary Process”, and Grant-in-Aid for Scientific Research on Innovative Areas from the Japanese Ministry of Education, Sports, Culture, Science, and Technology.

Author information

Authors and Affiliations

  1. Department of Chemistry, Tokyo Metropolitan University, Tokyo, Japan

    Yutaka Ito

  2. Advanced Science Institute, RIKEN, Saitama, Japan

    Tsutomu Mikawa

  3. School of Life Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK

    Brian O. Smith

Authors
  1. Yutaka Ito
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  2. Tsutomu Mikawa
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  3. Brian O. Smith
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Corresponding author

Correspondence to Yutaka Ito .

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Editors and Affiliations

  1. Department of Molecular Medicine, College of Medicine, University of South Florida, Bruce B. Downs Blvd 12901, Tampa, 33612, Florida, USA

    Vladimir N. Uversky

  2. Dept. Biochemistry & Molecular Biology, Center for Computational Biology &, Indiana University, West 10th Street 410, Indianapolis, 46202, Indiana, USA

    A. Keith Dunker

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Ito, Y., Mikawa, T., Smith, B.O. (2012). In-Cell NMR of Intrinsically Disordered Proteins in Prokaryotic Cells. In: Uversky, V., Dunker, A. (eds) Intrinsically Disordered Protein Analysis. Methods in Molecular Biology, vol 895. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-927-3_2

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  • DOI: https://doi.org/10.1007/978-1-61779-927-3_2

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-926-6

  • Online ISBN: 978-1-61779-927-3

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