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Journal of Chemical Biology

, Volume 5, Issue 3, pp 119–123 | Cite as

In situ synthesis of fluorescent membrane lipids (ceramides) using click chemistry

  • María Garrido
  • José Luis Abad
  • Alicia Alonso
  • Félix M. Goñi
  • Antonio Delgado
  • L.-Ruth MontesEmail author
Note

Abstract

Ceramide analogues containing azide groups either in the polar head or in the hydrocarbon chains are non-fluorescent. When incorporated into phospholipid bilayers, they can react in situ with a non-fluorescent 1,8-naphthalimide using click chemistry giving rise to fluorescent ceramide derivatives emitting at ≈440 nm. When incorporated into giant unilamellar vesicles, two-photon excitation at 760 nm allows visualization of the ceramide-containing bilayers. This kind of method may be of general applicability in the study of model and cell membranes.

Keywords

Ceramide GUV Fluorescence Click chemistry 

Abbreviations

DMSO

Dimethylsulfoxide

NMR

Nuclear magnetic resonance

HRMS

High-resolution mass spectrometry

ePC

Egg phosphatidylcholine

Notes

Acknowledgments

This work was supported in part by Spanish Ministerio de Ciencia e Innovación grants BFU2011-28566 (AA), SAF 2011-22444 (JLA/AD) and BFU 2007-62062 (FMG), and by Generalitat de Catalunya grant SGR 2009-1072 (JLA/AD). LRM is grateful to Consejo Superior de Investigaciones Científicas for a JAEdoc fellowship and MG for a JAEpredoc fellowship.

Supplementary material

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • María Garrido
    • 1
  • José Luis Abad
    • 1
  • Alicia Alonso
    • 2
  • Félix M. Goñi
    • 2
  • Antonio Delgado
    • 1
    • 3
  • L.-Ruth Montes
    • 2
    Email author
  1. 1.Consejo Superior de Investigaciones Científicas (CSIC), Research Unit on Bioactive Molecules (RUBAM)Institut de Química Avançada de Catalunya (IQAC-CSIC)BarcelonaSpain
  2. 2.Unidad de Biofísica (CSIC-UPV/EHU), and Departamento de BioquímicaUniversidad del País VascoBilbaoSpain
  3. 3.Facultad de Farmacia, Unidad de Química Farmacéutica (Unidad Asociada al CSIC)Universidad de Barcelona (UB)BarcelonaSpain

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