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Dynamics-Function Analysis in Catalytic RNA Using NMR Spin Relaxation and Conformationally Restricted Nucleotides

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Ribozymes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2167))

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Abstract

A full understanding of biomolecular function requires an analysis of both the dynamic properties of the system of interest and the identification of those dynamics that are required for function. We describe NMR methods based on metabolically directed specific isotope labeling for the identification of molecular disorder and/or conformational transitions on the RNA backbone ribose groups. These analyses are complemented by the use of synthetic covalently modified nucleotides constrained to a single sugar pucker, which allow functional assessment of dynamics by selectively removing a minor conformer identified by NMR from the structural ensemble.

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Acknowledgments

The authors are grateful to Dr. Victor Marquez and Dr. Ramon Eritja for helpful discussions and the U.S. National Science Foundation (MCB-1413356 to C.G.H.) and the Spanish Ministry of Economy (MINECO) (CTQ2017-84415-R to Ramon Eritja) for funding support.

Author information

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, USA

    Charles G. Hoogstraten, Neil A. White & Minako Sumita

  2. Institute for Advanced Chemistry of Catalonia (IQAC), Spanish Council for Scientific Research (CSIC), Barcelona, Spain

    Montserrat Terrazas & Anna Aviñó

  3. Joint IRB-BSC Program in Computational Biology, The Barcelona Institute of Science and Technology (BIST), Barcelona, Spain

    Montserrat Terrazas

  4. Networking Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Barcelona, Spain

    Anna Aviñó

  5. Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT, USA

    Neil A. White

  6. Department of Chemistry, Southern Illinois University Edwardsville, Edwardsville, IL, USA

    Minako Sumita

Authors
  1. Charles G. Hoogstraten
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  2. Montserrat Terrazas
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  3. Anna Aviñó
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  4. Neil A. White
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  5. Minako Sumita
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Corresponding author

Correspondence to Charles G. Hoogstraten .

Editor information

Editors and Affiliations

  1. Department of Microbiology and Immunology, Lady Davis Institute for Medical Research, Montréal, QC, Canada

    Robert J Scarborough

  2. Department of Microbiology and Immunology, Lady Davis Institute for Medical Research, Montréal, QC, Canada

    Anne Gatignol

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Hoogstraten, C.G., Terrazas, M., Aviñó, A., White, N.A., Sumita, M. (2021). Dynamics-Function Analysis in Catalytic RNA Using NMR Spin Relaxation and Conformationally Restricted Nucleotides. In: Scarborough, R.J., Gatignol, A. (eds) Ribozymes. Methods in Molecular Biology, vol 2167. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0716-9_11

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  • DOI: https://doi.org/10.1007/978-1-0716-0716-9_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0715-2

  • Online ISBN: 978-1-0716-0716-9

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