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Please use this identifier to cite or link to this item: http://hdl.handle.net/10481/44744

Title: Fluorogenic Substrates for In Situ Monitoring of Caspase-3 Activity in Live Cells
Authors: Pérez-López, Ana M.
Soria-Gila, M. Lourdes
Marsden, Emma R.
Lilienkampf, Annamaria
Bradley, Mark
Issue Date: 2016
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.
Sponsorship: This work was supported by the Ramon Areces and Caja Madrid Foundations to AMPL and Spanish Ministry of Economy and Competitiveness to MLSG (graduate student fellowships FPI BES-2010-030257 and EEBB-I-13-07131).
Publisher: Public Library of Science (PLOS)
Keywords: Apoptosis
Fluorescence imaging
Flow cytometry
Cross reactivity
Peptide synthesis
Apoptotic signaling cascade
High performance liquid chromatography
URI: http://hdl.handle.net/10481/44744
ISSN: 1932-6203
Rights : Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License
Citation: Pérez-López, A.M.; et al. Fluorogenic Substrates for In Situ Monitoring of Caspase-3 Activity in Live Cells. Plos One, 11(5): e0153209 (2016). [http://hdl.handle.net/10481/44744]
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