A novel quenched fluorescent activity-based probe reveals caspase-3 activity in the endoplasmic reticulum during apoptosis

The caspases are a family of cysteine proteases that are key regulators of apoptosis and their activity may thus serve as a good marker to monitor cell death. We have developed a quenched fluorescent activity-based probe (qABP) that is selective for caspase-3 activity and emits a fluorescent signal...

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Bibliographic Details
Published in:Chemical science (Cambridge) 2016-01, Vol.7 (2), p.1322-1337
Main Authors: Shaulov-Rotem, Yulia, Merquiol, Emmanuelle, Weiss-Sadan, Tommy, Moshel, Ofra, Salpeter, Seth, Shabat, Doron, Kaschani, Farnusch, Kaiser, Markus, Blum, Galia
Format: Article
Language:English
Subjects:
Activation
Apoptosis
Cell death
Endoplasmic reticulum
Fluorescence
Imaging
Monitors
Quenching (cooling)
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Summary:The caspases are a family of cysteine proteases that are key regulators of apoptosis and their activity may thus serve as a good marker to monitor cell death. We have developed a quenched fluorescent activity-based probe (qABP) that is selective for caspase-3 activity and emits a fluorescent signal after covalently modifying its target. The probe has a wide range of potential applications, e.g. in real-time imaging, FACS analysis or biochemical quantification of caspase activity in intact cells. Application of the probe allowed us to monitor caspase-3 activation after chemotherapy-treatment and to distinguish between apoptosis sensitive and resistant cells. Moreover, it enabled real-time high-resolution visualization of active caspase-3 during apoptosis. This led to the surprising finding that in cancerous cells active caspase-3 is not only found at the familiar cellular locations but also in mitochondria and the endoplasmic reticulum. Thus, our novel covalent probe allows high spatial and temporal resolution imaging of caspase-3 activation and may thus be used as an effective tool to study molecular mechanisms of programmed cell death in healthy and disease states. A selective quenched activity-based probe detects caspase-3 activity in the endoplasmic reticulum of cancerous cells during apoptosis.
ISSN:2041-6520
2041-6539
DOI:10.1039/c5sc03207e