TripleFRET measurements in flow cytometry

被引：14

作者：

Fabian, Akos ^{[1
]}

Horvath, Gabor ^{[2
]}

Vamosi, Gyoergy ^{[1
]}

Vereb, Gyoergy ^{[1
,3
]}

Szoellosi, Janos ^{[1
,3
]}

机构：

[1] Univ Debrecen, Dept Biophys & Cell Biol, Res Ctr Mol Med, Med & Hlth Sci Ctr, H-4012 Debrecen, Hungary

[2] Univ Bonn, Inst Innate Immun, Univ Hosp, Bonn, Germany

[3] Univ Debrecen, MTA Cell Biol & Signaling Res Grp, Res Ctr Mol Med, Med & Hlth Sci Ctr, H-4012 Debrecen, Hungary

来源：

CYTOMETRY PART A | 2013年 / 83A卷 / 04期

关键词：

Forster (fluorescence) resonance energy transfer; tripleFRET; relay transfer; trimeric complexes; RESONANCE ENERGY-TRANSFER; BY-CELL BASIS; LIVING CELLS; FRET MICROSCOPY; IMAGING MICROSCOPY; MOLECULAR-INTERACTIONS; SPECTROSCOPIC RULER; PROTEIN-INTERACTION; TRANSFER EFFICIENCY; SINGLE MOLECULES;

D O I：

10.1002/cyto.a.22267

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A frequently used method for viewing protein interactions and conformation, Forster (fluorescence) resonance energy transfer (FRET), has traditionally been restricted to two fluorophores. Lately, several methods have been introduced to expand FRET methods to three species. We present a method that allows the determination of FRET efficiency in three-dye systems on a flow cytometer. TripleFRET accurately reproduces energy transfer efficiency values measured in two-dye systems, and it can indicate the presence of trimeric complexes, which is not possible with conventional FRET methods. We also discuss the interpretation of energy transfer values obtained with tripleFRET in relation to spatial distribution of labeled molecules, specifically addressing the limitations of using total energy transfer to determine molecular distance. (c) 2013 International Society for Advancement of Cytometry

引用

页码：375 / 385

页数：11

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