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Bacterial Expression and In Vitro Refolding of Limited Fragments of the Notch Receptor and Its Ligands

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Book cover Notch Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1187))

Abstract

Prokaryotic expression of limited fragments of the Notch receptor and its ligands followed by in vitro refolding has been used for the production of the significant amounts of protein required for structure determination by X-ray crystallography or nuclear magnetic resonance spectroscopy. As an illustration of the protocol for the production of these EGF-containing constructs we have focused on a limited fragment of human Notch 1 that contains three calcium-binding EGF domains, hNotch-111–13. Following characterization by the methods described here, this construct has been shown to be functionally competent in a range of assays and the structure has been solved by X-ray crystallography and NMR.

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Acknowledgements

This work was supported by Cancer Research UK, the Wellcome Trust, and the EPA Cephalosporin Trust.

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Authors and Affiliations

  1. Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK

    Pat Whiteman, Christina Redfield & Penny A. Handford

Authors
  1. Pat Whiteman
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  2. Christina Redfield
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  3. Penny A. Handford
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Corresponding author

Correspondence to Penny A. Handford .

Editor information

Editors and Affiliations

  1. Dept.of Molecular and Human Genetics, Program in Developmental Biology, Dept. of Neuroscience, Jan and Dan Duncan Neurological Research Institute at Texas Children's Hospital, Howard Hughes Medical Institute, Baylor College of Medicine, Houston, Texas, USA

    Hugo J. Bellen

  2. Dept. of Molecular and Human Genetics, Program in Developmental Biology, Jan and Dan Duncan Neurological Research Institute at Texas Children's Hospital, Baylor College of Medicine, Houston, Texas, USA

    Shinya Yamamoto

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Whiteman, P., Redfield, C., Handford, P.A. (2014). Bacterial Expression and In Vitro Refolding of Limited Fragments of the Notch Receptor and Its Ligands. In: Bellen, H., Yamamoto, S. (eds) Notch Signaling. Methods in Molecular Biology, vol 1187. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1139-4_15

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  • DOI: https://doi.org/10.1007/978-1-4939-1139-4_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1138-7

  • Online ISBN: 978-1-4939-1139-4

  • eBook Packages: Springer Protocols

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