Department of Molecular Biophysics, Universiteit Utrecht, Princetonplein 1, 3584 CC Utrecht, The Netherlands.
Chemphyschem. 2011 Feb 25;12(3):475-83. doi: 10.1002/cphc.201000801. Epub 2010 Dec 22.
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.
同型Förster 共振能量转移（Homo-FRET）是指相同荧光基团之间的Förster 共振能量转移，可以通过观察其对荧光各向异性的影响来方便地测量。本综述旨在总结荧光各向异性成像技术在研究相同蛋白质和脂质聚集方面的可能性。同型 FRET 成像能够确定荧光团之间的距离。此外，它还可以用于定量测定簇的大小以及簇大小分布。同型 FRET 信号的解释很复杂，因为荧光团的相互取向以及每个簇中的荧光团数量都会以相似的方式影响荧光各向异性。荧光探针的特性非常重要。考虑到这些特性，对于在蛋白质和脂质聚集研究中正确解释同型 FRET 信号至关重要。这可以通过脂质筏标记物 GPI 和 K-ras 的聚集以及质膜中 EGF 受体聚集的研究来举例说明。
