Unprecedented Sensitivity in a Probe for Monitoring CathepsinB: Chemiluminescence Microscopy Cell-Imaging of a Natively Expressed Enzyme

Until recently, chemiluminescence cell images could only be obtained using luciferase-activated probes. Moreover, chemiluminescence microscopy cell-imaging has not been demonstrated for natively expressed enzymes like cathepsin B. Herein, we describe the design, synthesis, and evaluation of the firs...

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Bibliographic Details
Published inAngewandte Chemie International Edition Vol. 56; no. 49; pp. 15633 - 15638
Main Authors Roth-Konforti, Michal E., Bauer, Christoph R., Shabat, Doron
Format Journal Article
LanguageEnglish
Published WEINHEIM Wiley 04.12.2017
Subjects
Chemistry
Chemistry, Multidisciplinary
Physical Sciences
Science & Technology
CATALYZED CHEMILUMINESCENCE
dioxetanes
SUBSTITUTED DIOXETANE
cathepsin B
chemiluminescence
DOXORUBICIN
RELEASE
MODEL
1,2-DIOXETANES
live-cell imaging
molecular probes
PROTEASE ACTIVITY
BIOLUMINESCENCE
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Summary:Until recently, chemiluminescence cell images could only be obtained using luciferase-activated probes. Moreover, chemiluminescence microscopy cell-imaging has not been demonstrated for natively expressed enzymes like cathepsin B. Herein, we describe the design, synthesis, and evaluation of the first chemiluminescence probe for the detection and imaging of cathepsin B. The probe activation mechanism relies on the release of a dioxetane intermediate, which undergoes chemiexcitation to emit green light with high efficiency under physiological conditions. Using the probe, we obtained clear images of cancerous leukemia and colon cells. This is the first demonstration of chemiluminescence cell images obtained by a probe for a natively expressed endogenous enzyme. We anticipate that the concept presented in this study will be broadly used to develop analogous probes for other important proteases relevant to biomolecular processes.
ISSN:1433-7851
1521-3773
DOI:10.1002/anie.201709347