Thiol-reactive, luminescent Europium chelates: luminescence probes for resonance energy transfer distance measurements in biomolecules

Analytical Biochemistry
E Heyduk, T Heyduk

Abstract

Lanthanide chelates have recently been shown to be extremely promising luminescence probes for distance measurements in biomolecules using luminescence resonance energy transfer measurements [P. R. Selvin, T. M. Rana, and J. E. Hearst (1994) J. Am. Chem. Soc. 116, 6029-6030; P. R. Selvin, and J. E. Hearst (1994) Proc. Natl. Acad. Sci. USA 91, 10024-10028]. In this work we describe simple procedures for preparing highly fluorescent thiol-reactive europium chelates. These new compounds contain a uv-absorbing coumarin group which sensitizes europium emission, diethylenetriaminepentaacetic acid or triethylenetetraaminehexaacetic acid groups which provide europium chelating function, and a pyridyl disulfide group which allows specific modification of thiol groups. These reagents can be used to label proteins at Cys residues or synthetic oligonucleotides which contain thiol groups. Modification can be reversed easily by treatment with a reducing agent (dithiothreitol). Luminescence energy transfer between these new chelates and CY5 fluorochrome attached to the opposite ends of 15-bp double-stranded DNA was measured to test their usefulness for distance measurements in macromolecules. The distance measured between the chelate (donor) ...Continue Reading

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Citations

May 22, 2008·Analytical Chemistry·Ewa HeydukTomasz Heyduk
Jul 1, 2009·Analytical Chemistry·Ling Tian, Tomasz Heyduk
Jan 21, 2010·Bioconjugate Chemistry·Lev N KrasnoperovArkady Mustaev
Feb 24, 2006·Proceedings of the National Academy of Sciences of the United States of America·Steven E MansoorDavid L Farrens
Jul 9, 2013·Journal of Photochemistry and Photobiology. A, Chemistry·Shyamala PillaiArkady Mustaev
Jul 9, 2013·Journal of Photochemistry and Photobiology. A, Chemistry·Laura WirpszaArkady Mustaev
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Sep 29, 2009·Analytical Biochemistry·Ewa Heyduk, Tomasz Heyduk
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Feb 1, 2002·Nature Biotechnology·Tomasz Heyduk, Ewa Heyduk
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Nov 2, 2013·The Journal of Biological Chemistry·Sergio Barranco-MedinaEnrico Di Cera
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Apr 30, 2005·Analytical Chemistry·Ewa Heyduk, Tomasz Heyduk
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Feb 14, 2004·Analytical Chemistry·Eric Knoll, Tomasz Heyduk

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