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Bioluminescent properties of obelin and aequorin with novel coelenterazine analogues

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Abstract

The main analytical use of Ca2+-regulated photoproteins from luminous coelenterates is for real-time non-invasive visualization of intracellular calcium concentration ([Ca2+]i) dynamics in cells and whole organisms. A limitation of this approach for in vivo deep tissue imaging is the fact that blue light emitted by the photoprotein is highly absorbed by tissue. Seven novel coelenterazine analogues were synthesized and their effects on the bioluminescent properties of recombinant obelin from Obelia longissima and aequorin from Aequorea victoria were evaluated. Only analogues having electron-donating groups (m-OCH3 and m-OH) on the C6 phenol moiety or an extended resonance system at the C8 position (1-naphthyl and α-styryl analogues) showed a significant red shift of light emission. Of these, only the α-styryl analogue displayed a sufficiently high light intensity to allow eventual tissue penetration. The possible suitability of this compound for in vivo assays was corroborated by studies with aequorin which allowed the monitoring of [Ca2+]i dynamics in cultured CHO cells and in hippocampal brain slices. Thus, the α-styryl coelenterazine analogue might be potentially useful for non-invasive, in vivo bioluminescence imaging in deep tissues of small animals.

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Abbreviations

BRET:

Bioluminescence Förster resonance energy transfer

CHO cells:

Chinese hamster ovary cells

CTZ:

Coelenterazine

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Acknowledgements

R.G. acknowledges the ICSN for a fellowship. We are grateful for the ANR grant to P.B. and a CNRS Physics, Chemistry and Biology interface grant to R.H.D. and P.B.; N.P.M, L.P.B., and E.S.V. acknowledge the RFBR grant 12-04-00131 and the Program of the Government of Russian Federation “Measures to attract leading scientists to Russian educational institutions” (grant 11.G34.31.0058). P.B. and A.J.B. are indebted to Eric Karplus from Science Wares Inc. for helping with single-photon imaging software.

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Authors and Affiliations

  1. Institut de Chimie des Substances Naturelles, UPR 2301, Centre National de la Recherche Scientifique, Avenue de la Terrasse, 91198, Gif-sur-Yvette, France

    Ronan Gealageas & Robert H. Dodd

  2. Photobiology Laboratory, Institute of Biophysics, Russian Academy of Sciences, Siberian Branch, Krasnoyarsk, 660036, Russia

    Natalia P. Malikova, Ludmila P. Burakova & Eugene S. Vysotski

  3. Laboratory of Bioluminescent Biotechnologies, Institute of Fundamental Biology and Biotechnology, Siberian Federal University, Krasnoyarsk, 660041, Russia

    Natalia P. Malikova, Ludmila P. Burakova & Eugene S. Vysotski

  4. Institut des Neurosciences Alfred Fessard, UPR 3294, Centre National de la Recherche Scientifique, Avenue de la Terrasse, 91198, Gif-sur-Yvette, France

    Sandrine Picaud, Aren J. Borgdorff & Philippe Brûlet

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  1. Ronan Gealageas
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  2. Natalia P. Malikova
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  3. Sandrine Picaud
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  4. Aren J. Borgdorff
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  5. Ludmila P. Burakova
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  6. Philippe Brûlet
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  7. Eugene S. Vysotski
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  8. Robert H. Dodd
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Corresponding author

Correspondence to Robert H. Dodd.

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Ronan Gealageas, Natalia P. Malikova, and Sandrine Picaud contributed equally to this work.

Philippe Brûlet is deceased.

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Gealageas, R., Malikova, N.P., Picaud, S. et al. Bioluminescent properties of obelin and aequorin with novel coelenterazine analogues. Anal Bioanal Chem 406, 2695–2707 (2014). https://doi.org/10.1007/s00216-014-7656-4

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  • DOI: https://doi.org/10.1007/s00216-014-7656-4

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