title: Homo-FRET imaging as a tool to quantify protein and lipid clustering.
abstract: Homo-FRET, Förster resonance energy transfer between identical fluorophores, can be conveniently measured by observing its effect on the fluorescence anisotropy. This review aims to summarize the possibilities of fluorescence anisotropy imaging techniques to investigate clustering of identical proteins and lipids. Homo-FRET imaging has the ability to determine distances between fluorophores. In addition it can be employed to quantify cluster sizes as well as cluster size distributions. The interpretation of homo-FRET signals is complicated by the fact that both the mutual orientations of the fluorophores and the number of fluorophores per cluster affect the fluorescence anisotropy in a similar way. The properties of the fluorescence probes are very important. Taking these properties into account is critical for the correct interpretation of homo-FRET signals in protein- and lipid-clustering studies. This is be exemplified by studies on the clustering of the lipid raft markers GPI and K-ras, as well as for EGF receptor clustering in the plasma membrane.
authors.affiliation: Department of Molecular Biophysics, Universiteit Utrecht, Princetonplein 1, 3584 CC Utrecht, The Netherlands.
authors.forename: Arjen N
authors.initials: AN
authors.lastname: Bader
authors.forename: Sandra
authors.initials: S
authors.lastname: Hoetzl
authors.forename: Erik G
authors.initials: EG
authors.lastname: Hofman
authors.forename: Jarno
authors.initials: J
authors.lastname: Voortman
authors.forename: Paul M P
authors.initials: PM
authors.lastname: van Bergen en Henegouwen
authors.forename: Gerrit
authors.initials: G
authors.lastname: van Meer
authors.forename: Hans C
authors.initials: HC
authors.lastname: Gerritsen
mesh_terms: D016000:Cluster Analysis; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D008055:Lipids / Q000032:analysis*; D008856:Microscopy, Fluorescence; D011506:Proteins / Q000032:analysis*
chemical_list: D005456:Fluorescent Dyes; D008055:Lipids; D011506:Proteins