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

A1 Journal article (refereed)


Internal Authors/Editors


Publication Details

List of Authors: Najumudeen AK, Guzmán C, Posada IMD, Abankwa D
Publisher: Humana Press, Inc.
Place: Springer New York
Publication year: 2015
Journal: Methods in Molecular Biology
Volume number: 1298
Start page: 29
End page: 45
eISSN: 1940-6029


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.





Last updated on 2019-17-11 at 04:11