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

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.

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Abbreviations

DMSO:

Dimethylsulfoxide

NMR:

Nuclear magnetic resonance

HRMS:

High-resolution mass spectrometry

ePC:

Egg phosphatidylcholine

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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.

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Correspondence to L.-Ruth Montes.

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Garrido, M., Abad, J.L., Alonso, A. et al. In situ synthesis of fluorescent membrane lipids (ceramides) using click chemistry. J Chem Biol 5, 119–123 (2012). https://doi.org/10.1007/s12154-012-0075-0

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Keywords

  • Ceramide
  • GUV
  • Fluorescence
  • Click chemistry