Probing phospholipase A2 with fluorescent phospholipid substrates

Oliver Wichmann, Michael H. Gelb, Carsten Schultz

Research output: Contribution to journalArticlepeer-review

36 Scopus citations


The Foerster resonance energy transfer-based sensor, PENN, measures intracellular phospholipase A2 (PLA2) activity in living cells and small organisms. In an attempt to modify the probe for the detection of particular isoforms, we altered the sn-2 fatty acid in such a way that either one or three of the Z double bonds in arachidonic acid were present in the sensor molecule. Arachidonic-acid-mimicking fatty acids were prepared by copper-mediated coupling reactions. Probes with a single double bond in the 5-position exhibited favorable substrate properties for secretory PLA 2s. In vitro experiments with the novel unsaturated doubly labeled phosphatidylethanolamine derivatives showed preferred cleavage of the sensor PENN2 (one double bond) by the physiologically important group V sPLA 2, while the O-methyl-derivative PMNN2 was accepted best by the isoform from hog pancreas. For experiments in living cells, we demonstrated that bioactivation via S-acetylthioethyi (SATE) groups is essential for probe performance. Surprisingly, membrane-permeant versions of the new sensors that contained double bonds, PENN2 and PENN3, were only cleaved to a minor extent in HeLa cells while the saturated form, PENN, was well accepted.

Original languageEnglish (US)
Pages (from-to)1555-1569
Number of pages15
Issue number13
StatePublished - Sep 3 2007
Externally publishedYes


  • Alkynes
  • Bioorganic chemistry
  • FRET
  • Fluorescent probes
  • Phospholipase
  • Phospholipids

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry


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