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Cell Surface Biotinylation of Receptor Tyrosine Kinases to Investigate Intracellular Trafficking

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Book cover Receptor Tyrosine Kinases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1233))

Abstract

Cell surface biotinylation is a biochemical approach to covalently bind membrane-impermeable biotin to the extracellular domain of membrane proteins, such as receptor tyrosine kinases (RTKs). Subsequent to ligand incubation periods, activated biotinylated receptors may internalize from the cell surface into early endosomes and then travel through intracellular compartments to either recycle back to the membrane or degrade in lysosomes. The biotin-labeled proteins may be detected through affinity purification with streptavidin agarose resins. This chapter describes methods for cell surface biotinylation to assess RTK trafficking steps.

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Acknowledgements

This work was supported by a grant from the Cancer Research Society and Carcinoid NeuroEndocrine Tumour Society Canada (L.M. Mulligan). M.J.F. Crupi is the recipient of an Ontario Graduate Scholarship doctoral award.

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Authors and Affiliations

  1. Division of Cancer Biology and Genetics, Cancer Research Institute, Queen’s University, Botterell Hall Rm A315, Kingston, ON, Canada, K7L 3N6

    Mathieu J. F. Crupi, Douglas S. Richardson & Lois M. Mulligan

  2. Department of Pathology & Molecular Medicine, Queen’s University, Kingston, ON, Canada

    Mathieu J. F. Crupi, Douglas S. Richardson & Lois M. Mulligan

  3. Center for Biological Imaging, Harvard University, Cambridge, MA, 02138, USA

    Douglas S. Richardson

Authors
  1. Mathieu J. F. Crupi
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  2. Douglas S. Richardson
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  3. Lois M. Mulligan
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Corresponding author

Correspondence to Lois M. Mulligan .

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Editors and Affiliations

  1. Necker Hospital, Paris, France

    Serena Germano

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© 2015 Springer Science+Business Media New York

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Crupi, M.J.F., Richardson, D.S., Mulligan, L.M. (2015). Cell Surface Biotinylation of Receptor Tyrosine Kinases to Investigate Intracellular Trafficking. In: Germano, S. (eds) Receptor Tyrosine Kinases. Methods in Molecular Biology, vol 1233. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1789-1_9

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  • DOI: https://doi.org/10.1007/978-1-4939-1789-1_9

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1788-4

  • Online ISBN: 978-1-4939-1789-1

  • eBook Packages: Springer Protocols

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