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

A1 Originalartikel i en vetenskaplig tidskrift (referentgranskad)


Interna författare/redaktörer


Publikationens författare: Najumudeen AK, Guzmán C, Posada IMD, Abankwa D
Förläggare: Humana Press, Inc.
Förlagsort: Springer New York
Publiceringsår: 2015
Tidskrift: Methods in Molecular Biology
Volym: 1298
Artikelns första sida, sidnummer: 29
Artikelns sista sida, sidnummer: 45
eISSN: 1940-6029


Abstrakt




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.





Senast uppdaterad 2019-24-06 vid 04:23