Abstract
Proteases are one of the most abundant classes of enzymes and are involved in a plethora of biological processes in many cellular compartments, including the mitochondria. To understand the role of proteases is essential to determine their substrate repertoire, preferably in an in vivo setting. In this chapter we describe general guidelines to analyze protease activity using several strategies, from in-gel analysis to mass spectrometry mapping of the cleavage site(s) and fluorogenic probes that can easily be used in vivo. To exemplify this flowchart, we used the recently characterized organellar oligopeptidase of Arabidopsis (Arabidopsis thaliana), an enzyme that takes part in degradation of short peptides within mitochondria and chloroplasts.
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Acknowledgement
This work was supported by grants from the Swedish Research Council to EG.
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Teixeira, P.F., Branca, R.M., Kmiec, B., Glaser, E. (2015). A Flowchart to Analyze Protease Activity in Plant Mitochondria. In: Whelan, J., Murcha, M. (eds) Plant Mitochondria. Methods in Molecular Biology, vol 1305. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2639-8_8
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DOI: https://doi.org/10.1007/978-1-4939-2639-8_8
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2638-1
Online ISBN: 978-1-4939-2639-8
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