Üstün, Suayib
- Department of Plant Biology, Swedish University of Agricultural Sciences
Abstract
The fine-tuned balance of protein level, conformation and location within the cell is vital for the dynamic changes required for a cell to respond to a given stimulus. This requires the regulated turnover of damaged or short-lived proteins through the ubiquitin proteasome system (UPS). Thus, the protease activity of the proteasome is adjusted to meet the current demands of protein degradation via the UPS within the cell. We describe the adaptation of an intramolecular quenched fluorescence assay utilizing substrate-mimic peptides for the measurement of proteasome activity in total plant extracts. The peptide substrates contain donor-quencher pairs that flank the scissile bond. Following cleavage, the increase in dequenched donor emission of the product is subsequently measured over time and used to calculate the relative proteasome activity.
Keywords
Proteasome; Immunity; Plant defense; Targeted proteolysis; Fluorogenic substrate
Published in
Bio-protocol
2017, volume: 7, number: 17, article number: e2532
Publisher: BIO-PROTOCOL
SLU Authors
UKÄ Subject classification
Botany
Publication identifier
- DOI: https://doi.org/10.21769/BioProtoc.2532
Permanent link to this page (URI)
https://res.slu.se/id/publ/105574