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Rab GTPases pp 29–45Cite as

Rab-NANOPS: FRET Biosensors for Rab Membrane Nanoclustering and Prenylation Detection in Mammalian Cells

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1298))

Abstract

Rab proteins constitute the largest subfamily of Ras-like small GTPases. They are central to vesicular transport and organelle definition in eukaryotic cells. Unlike their Ras counterparts, they are not a hallmark of cancer. However, a number of diseases, including cancer, show a misregulation of Rab protein activity. As for all membrane-anchored signaling proteins, correct membrane organization is critical for Rabs to operate. In this chapter, we provide a detailed protocol for the use of a flow cytometry-based Fluorescence Resonance Energy Transfer (FRET)-biosensors assay, which allows to detect changes in membrane anchorage, subcellular distribution, and of the nanoscale organization of Rab-GTPases in mammalian cell lines. This assay is high-throughput amenable and can therefore be utilized in chemical-genomic and drug discovery efforts.

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Authors and Affiliations

  1. Turku Centre for Biotechnology, Åbo Akademi University, Tykistökatu 6B, 20520, Turku, Finland

    Arafath Kaja Najumudeen, Camilo Guzmán, Itziar M. D. Posada & Daniel Abankwa

Authors
  1. Arafath Kaja Najumudeen
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  2. Camilo Guzmán
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  3. Itziar M. D. Posada
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  4. Daniel Abankwa
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Corresponding author

Correspondence to Daniel Abankwa .

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Editors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Peggy and Charles Stephenson Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA

    Guangpu Li

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Najumudeen, A.K., Guzmán, C., Posada, I.M.D., Abankwa, D. (2015). Rab-NANOPS: FRET Biosensors for Rab Membrane Nanoclustering and Prenylation Detection in Mammalian Cells. In: Li, G. (eds) Rab GTPases. Methods in Molecular Biology, vol 1298. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2569-8_3

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  • DOI: https://doi.org/10.1007/978-1-4939-2569-8_3

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2568-1

  • Online ISBN: 978-1-4939-2569-8

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