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Biochemical Responses to Chemoattractants in Dictyostelium: Ligand-Receptor Interactions and Downstream Kinase Activation

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Chemotaxis

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 571))

Summary

Dictyostelium discoideum is one of the most facile eukaryotic systems for the study of chemotactic response to secreted chemical ligands. Dictyostelium grow as individual cells, using bacteria and fungi as primary nutrient sources; during growth, Dictyostelium moves directionally toward folate, a bacterial byproduct. Upon nutrient depletion Dictyostelium initiates a multicellular development program characterized by the production and secretion of cAMP. Cell surface receptors specifically recognize extracellular cAMP, which serves as both a morphogen to promote development and a chemoattractant to organize multicellularity. We discuss several approaches for the study of ligand–receptor interaction, with focus on affinity class determination and quantification of ligand binding sites (i.e., receptors) per cell. We further present examples for the application of biochemical assays to characterize the ligand-induced kinase activation of PI3K, GSK3, and ERK2.

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Acknowledgments

This work was supported by the Intramural Research Program of the National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health. We are indebted to the helpful comments and interest of Vanessa McMains and Jonathon Buggey.

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Authors and Affiliations

  1. Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, NIH, Bethesda, MD, USA

    Xin-Hua Liao & Alan R. Kimmel

Authors
  1. Xin-Hua Liao
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  2. Alan R. Kimmel
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Editor information

Editors and Affiliations

  1. Infectious Diseases (NIAID), National Institute of Allergy and, Parklawn Dr. 12441, Rockville, 20852, U.S.A.

    Tian Jin

  2. National Institute on Alcohol Abuse &, National Institute of Health, Fisher Lane 5635, Bethesda, 20892-9304, U.S.A.

    Dale Hereld

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© 2009 Humana Press

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Liao, XH., Kimmel, A.R. (2009). Biochemical Responses to Chemoattractants in Dictyostelium: Ligand-Receptor Interactions and Downstream Kinase Activation. In: Jin, T., Hereld, D. (eds) Chemotaxis. Methods in Molecular Biology™, vol 571. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-198-1_18

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  • DOI: https://doi.org/10.1007/978-1-60761-198-1_18

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-197-4

  • Online ISBN: 978-1-60761-198-1

  • eBook Packages: Springer Protocols

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