Fluorogenic Substrates for In Situ Monitoring of Caspase-3 Activity in Live Cells

Ana M. Perez-Lopez, M. Lourdes Soria-Gila, Emma R. Marsden, Annamaria Lilienkampf, Mark Bradley*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

The in situ detection of caspase-3 activity has applications in the imaging and monitoring of multiple pathologies, notably cancer. A series of cell penetrating FRET-based fluorogenic substrates were designed and synthesised for the detection of caspase-3 in live cells. A variety of modifications of the classical caspase-3 and caspase-7 substrate sequence Asp-Glu-Val-Asp were carried out in order to increase caspase-3 affinity and eliminate caspase-7 cross-reactivity. To allow cellular uptake and good solubility, the substrates were conjugated to a cationic peptoid. The most selective fluorogenic substrate 27, FAM-Ahx-Asp-Leu-Pro-Asp-Lys(MR)-Ahx, conjugated to the cell penetrating peptoid at the C-terminus, was able to detect and quantify caspase-3 activity in apoptotic cells without cross-reactivity by caspase-7.

Original languageEnglish
Article number0153209
Number of pages11
JournalPLoS ONE
Volume11
Issue number5
DOIs
Publication statusPublished - 11 May 2016

Keywords

  • ACTIVITY-BASED PROBES
  • UNNATURAL AMINO-ACIDS
  • PENETRATING PEPTIDES
  • APOPTOTIC PATHWAYS
  • CANCER
  • SPECIFICITY
  • CLEAVAGE
  • INHIBITORS
  • DISCOVERY
  • PROTEASES

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