Projects per year
The ubiquitin-proteasome system (UPS) is the predominant protein degradation machinery in eukaryotic cells. It is highly conserved among eukaryotes and essential for their survival. Through regulated proteolysis the UPS plays a key role in a myriad of cellular functions, including developmental and stress signaling, cell differentiation, and cell death. Attachment of a ubiquitin chain to a substrate can trigger its recruitment to the proteasome for proteolysis. To efficiently degrade substrates, however, the proteasome employs HECT-type ubiquitin ligases that can further remodel ubiquitin chains of proteasome-captured substrates. It is thought that this remodeling process is necessary to maintain substrate affinity for the proteasome and to completely translocate the substrate into the 20S proteolytic barrel. Here, we describe a protocol for purifying proteasomes and their associated accessory proteins and provide a practical way to detect proteasome-associated E3 ligase activity. This assay is reliable and efficient for assessing the ability of proteasomes to form ubiquitin conjugates and is applicable to a wide range of eukaryotic species.
|Title of host publication||Plant Proteostasis|
|Subtitle of host publication||Methods in Molecular Biology|
|Publication status||Published - 23 Nov 2022|
|Name||Methods in Molecular Biology|
- HECT-type ligases
- posttranslational modifications
- proteasome-associated ubiquitin E3 ligases
FingerprintDive into the research topics of 'Analysis of proteasome-associated ubiquitin ligase activity'. Together they form a unique fingerprint.
UbRegulate: Dynamic ubiquitin signalling coordinates transcriptional reprogramming in plant immunity
1/09/21 → 31/08/26
1/12/19 → 1/04/23
1/03/16 → 31/08/21