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Circular Dichroism and Fluorescence Spectroscopy to Study Protein Structure and Protein–Protein Interactions in Ethylene Signaling

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Ethylene Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1573))

Abstract

Circular dichroism (CD) spectroscopy is an invaluable technique to analyze secondary structure and functional folding of recombinant purified proteins. CD spectroscopy can also be applied to detect changes in protein secondary structure related to the pH or redox conditions found in different cellular compartments or to the interaction with other molecules. Another biophysical technique to monitor conformational changes and interaction with small molecule ligands or biological macromolecules is protein fluorescence spectroscopy making use of the aromatic amino acid tryptophan as a sensitive intrinsic fluorescent probe. Here, we describe the application of CD and tryptophan fluorescence spectroscopy to study soluble and membrane proteins of the ethylene signaling pathway.

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Acknowledgments

The authors would like to thank Dr. Silke Allekotte, Dr. Melanie Bisson-Ritter, and Dr. Elisa Classen for CD and fluorescence measurements on purified ETR1 and EIN2. The critical reading of the manuscript by Dr. Melanie Bisson-Ritter and Lena Müller is also greatly acknowledged. Support for work in the authors’ laboratory from grants from the Deutsche Forschungsgemeinschaft (GR1616-7, CRC590, GR1616-9, GR1616-10) is gratefully acknowledged.

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Authors and Affiliations

  1. Institute of Biochemical Plant Physiology, Heinrich-Heine University Düsseldorf, Universitätsstraße 1, D-40225, Düsseldorf, Germany

    Mareike Kessenbrock & Georg Groth

Authors
  1. Mareike Kessenbrock
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  2. Georg Groth
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Corresponding author

Correspondence to Georg Groth .

Editor information

Editors and Affiliations

  1. Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, Tennessee, USA

    Brad M. Binder

  2. Department of Biological Sciences, Dartmouth College, Hanover, New Hampshire, USA

    G. Eric Schaller

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Kessenbrock, M., Groth, G. (2017). Circular Dichroism and Fluorescence Spectroscopy to Study Protein Structure and Protein–Protein Interactions in Ethylene Signaling. In: Binder, B., Eric Schaller, G. (eds) Ethylene Signaling. Methods in Molecular Biology, vol 1573. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6854-1_12

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  • DOI: https://doi.org/10.1007/978-1-4939-6854-1_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6852-7

  • Online ISBN: 978-1-4939-6854-1

  • eBook Packages: Springer Protocols

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