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J Exp Bot. 2019 Feb 26. pii: erz076. doi: 10.1093/jxb/erz076. [Epub ahead of print]

Caught green-handed: methods for in vivo detection and visualization of protease activity.

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Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark, Ghent, Belgium.
VIB Center for Plant Systems Biology, Technologiepark, Ghent, Belgium.
The Plant Chemetics Laboratory, Department of Plant Sciences, University of Oxford, Oxford, UK.
Department of Biomolecular Medicine, Ghent University, Ghent, Belgium.
VIB Center for Medical Biotechnology, Ghent, Belgium.


Proteases are enzymes that cleave peptide bonds of other proteins. Their omnipresence and diverse activities make them important players in protein homeostasis and turnover of the total cell proteome as well as in signal transduction in plant stress response and development. To fully understand protease function, it is of paramount importance to assess when and where a specific protease is active. Here, we review the existing methods to detect in vivo protease activity by means of imaging chemical activity-based probes and genetically encoded sensors. We focus on the diverse fluorescent and luminescent sensors at the researcher's disposal and evaluate the potential of imaging techniques to deliver in vivo spatiotemporal detail of protease activity. We predict that in the coming years, revised techniques will help to elucidate plant protease activity, functions and hence expand the current status of the field.


in vivo imaging; activity-based probes; fluorescent; luminescent; plants; protease; proteolysis; reporter; sensor


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