Rab-NANOPS: FRET biosensors for Rab membrane nanoclustering and prenylation detection in mammalian cells

Arafath Kaja Najumudeen; Camilo Guzman; Posada IMD; Daniel Abankwa

doi:10.1007/978-1-4939-2569-8_3

Rab-NANOPS: FRET biosensors for Rab membrane nanoclustering and prenylation detection in mammalian cells

Arafath Kaja Najumudeen, Camilo Guzman, Posada IMD, Daniel Abankwa

Research output: Contribution to journal › Article › Scientific › peer-review

4 Citations (Scopus)

Abstract

Rab proteins constitute the largest subfamily of Ras-like small GTPases. They are central to vesicular trans- port 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, subcel- lular 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.

Original language	Undefined/Unknown
Pages (from-to)	29–45
Journal	Methods in Molecular Biology
Volume	1298
DOIs	https://doi.org/10.1007/978-1-4939-2569-8_3
Publication status	Published - 2015
MoE publication type	A1 Journal article-refereed

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10.1007/978-1-4939-2569-8_3

Cite this

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title = "Rab-NANOPS: FRET biosensors for Rab membrane nanoclustering and prenylation detection in mammalian cells",

abstract = "Rab proteins constitute the largest subfamily of Ras-like small GTPases. They are central to vesicular trans- port 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, subcel- lular 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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AU - Guzman, Camilo

AU - IMD, Posada

AU - Abankwa, Daniel

PY - 2015

Y1 - 2015

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AB - Rab proteins constitute the largest subfamily of Ras-like small GTPases. They are central to vesicular trans- port 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, subcel- lular 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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