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Innate DNA and RNA Recognition pp 27–35Cite as

Crystallization of Mouse RIG-I ATPase Domain: In Situ Proteolysis

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1169))

Abstract

RIG-I is a key pattern recognition receptor that recognizes cytoplasmic viral RNA. Upon ligand binding, it undergoes a conformational change that induces an active signaling conformation. However, the details of this conformational change remain elusive until high-resolution crystal structures of different functional conformations are available. X-ray crystallography is a powerful tool to study structure–function relationships, but crystallization is often the limiting step of the method. Here, we describe the in situ in-drop proteolysis of RIG-I that yielded crystals of the ATPase domain of mouse RIG-I suitable for structure determination.

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Acknowledgments

This work was funded by National Institutes of Health grant U19AI083025 and grants from the Deutsche Forschungsgemeinschaft (DFG HO2489/3 and SFB455) and financial support from Center for Integrated Protein Science Munich to KPH.

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Authors and Affiliations

  1. Department of Biochemistry at the Gene Center, Ludwig-Maximilians-University Munich, Feodor-Lynen-Strasse 25, 81377, Munich, Germany

    Filiz Civril & Karl-Peter Hopfner

  2. Graduate School for Quantivative Biosciences Munich, Ludwig-Maximilians-University Munich, Munich, Germany

    Filiz Civril & Karl-Peter Hopfner

Authors
  1. Filiz Civril
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  2. Karl-Peter Hopfner
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Corresponding author

Correspondence to Karl-Peter Hopfner .

Editor information

Editors and Affiliations

  1. Ludwig Maximilians Universität München, Medizinische Klinik und Poliklinik IV, München, Germany

    Hans-Joachim Anders

  2. Helmholtz Zentrum München German Researc, Institute for Diabetes and Regeneration, Munich, Germany

    Adriana Migliorini

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Civril, F., Hopfner, KP. (2014). Crystallization of Mouse RIG-I ATPase Domain: In Situ Proteolysis. In: Anders, HJ., Migliorini, A. (eds) Innate DNA and RNA Recognition. Methods in Molecular Biology, vol 1169. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0882-0_3

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  • DOI: https://doi.org/10.1007/978-1-4939-0882-0_3

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0881-3

  • Online ISBN: 978-1-4939-0882-0

  • eBook Packages: Springer Protocols

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