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Chemotaxis pp 63–77Cite as

Studying Chemoattractant Signal Transduction Dynamics in Dictyostelium by BRET

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1407))

Abstract

Understanding the dynamics of chemoattractant signaling is key to our understanding of the mechanisms underlying the directed migration of cells, including that of neutrophils to sites of infections and of cancer cells during metastasis. A model frequently used for deciphering chemoattractant signal transduction is the social amoeba Dictyostelium discoideum. However, the methods available to quantitatively measure chemotactic signaling are limited. Here, we describe a protocol to quantitatively study chemoattractant signal transduction in Dictyostelium by monitoring protein–protein interactions and conformational changes using Bioluminescence Resonance Energy Transfer (BRET).

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Acknowledgements

We are thankful to Michel Bouvier for providing BRET2 donor and acceptor cDNAs, to Chris Jenetopoulos for providing the Dictyostelium Gα2 and Gβ cDNAs, and to Billy Breton for constructive discussions and critical reading of the manuscript.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of Arizona, 1041 E. Lowell Street, Tucson, 85721-0088, AZ, USA

    A. F. M. Tariqul Islam, Branden M. Stepanski & Pascale G. Charest

Authors
  1. A. F. M. Tariqul Islam
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  2. Branden M. Stepanski
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  3. Pascale G. Charest
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Corresponding author

Correspondence to Pascale G. Charest .

Editor information

Editors and Affiliations

  1. Nat Inst of Allergy& Infectious Dis, NIH, Laboratory of Immunogenetics, Rockville, Maryland, USA

    Tian Jin

  2. National Institutes of Health, Nat Inst Alcohol Abuse & Alcoholism, Rockville, Maryland, USA

    Dale Hereld

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Islam, A.F.M.T., Stepanski, B.M., Charest, P.G. (2016). Studying Chemoattractant Signal Transduction Dynamics in Dictyostelium by BRET. In: Jin, T., Hereld, D. (eds) Chemotaxis. Methods in Molecular Biology, vol 1407. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3480-5_5

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  • DOI: https://doi.org/10.1007/978-1-4939-3480-5_5

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3478-2

  • Online ISBN: 978-1-4939-3480-5

  • eBook Packages: Springer Protocols

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