Homogeneous time resolved fluorescence resonance energy transfer using rare earth cryptates as a tool for probing molecular interactions in biology.
A homogeneous assay technology using time resolved fluorescence and fluorescence resonance energy transfer is described. A new class of fluorescent complexes, the cryptates, have been used as fluorescent donor with cross-linked allophycocyanin as acceptor. This new donor/acceptor shows an exceptionally high Förster distance R0 of 9 nm. This allows to build up a set of strategies to probe the interactions of biomolecules in biology, particularly for high throughput screening applications. In this article, we describe the basics of the technology and review applications developed for studying different key molecular interactions involved in cellular processes.
Bazin
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CIS Bio International, Bagnols sur Céze, France.
Préaudat
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Trinquet
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Mathis
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D002462:Cell Membrane / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry*; D007700:Kinetics; D008024:Ligands; D008956:Models, Chemical; D009942:Organometallic Compounds / Q000737:chemistry*; D010798:Phycocyanin / Q000737:chemistry; D011485:Protein Binding; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013816:Thermodynamics; D013997:Time Factors
D005456:Fluorescent Dyes; D008024:Ligands; D009942:Organometallic Compounds; C002497:allophycocyanin; D010798:Phycocyanin; C070734:europium(III) trisbipyridine cryptate