ǂa ǂdual-function cysteine protease with endo- and transpeptidase activity
Katarina Petra van Midden (Author), Melissa Mantz (Author), Marko Fonović (Author), Martin Gazvoda (Author), Jurij Svete (Author), Pitter F. Huesgen (Author), Renier A. L. van der Hoorn (Author), Marina Klemenčič (Author)

Abstract

Proteases, essential regulators of plant stress responses, remain enigmatic in their precise functional roles. By employing activity-based probes for real-time monitoring, this study aimed to delve into protease activities in Chlamydomonas reinhardtii exposed to oxidative stress induced by hydrogen peroxide. However, our work revealed that the activity-based probes strongly labelled three non-proteolytic proteins—PsbO, PsbP, and PsbQ—integral components of photosystem II's oxygen-evolving complex. Subsequent biochemical assays and mass spectrometry experiments revealed the involvement of CrCEP1, a previously uncharacterized papain-like cysteine protease, as the catalyst of this labelling reaction. Further experiments with recombinant CrCEP1 and PsbO proteins replicated the reaction in vitro. Our data unveiled that endopeptidase CrCEP1 also has transpeptidase activity, ligating probes and peptides to the N-termini of Psb proteins, thereby expanding the repertoire of its enzymatic activities. The hitherto unknown transpeptidase activity of CrCEP1, working in conjunction with its proteolytic activity, unveils putative complex and versatile roles for proteases in cellular processes during stress responses.

Keywords

proteoliza;sonde na osnovi aktivnosti;papainu podobna cisteinska proteaza;transpeptidacija;zelene alge;proteolysis;activity-based probes;papain-like cysteine protease;transpeptidation;green algae;

Data

Language: English
Year of publishing:
Typology: 1.01 - Original Scientific Article
Organization: UL FKKT - Faculty of Chemistry and Chemical Technology
UDC: 577.15
COBISS: 197196291 Link will open in a new window
ISSN: 0141-8130
Views: 62
Downloads: 17
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Other data

Secondary language: Slovenian
Secondary keywords: proteoliza;sonde na osnovi aktivnosti;papainu podobna cisteinska proteaza;transpeptidacija;zelene alge;
Type (COBISS): Article
Pages: str. 1-15
Volume: ǂVol. ǂ271
Issue: ǂpt. ǂ1, [article no.] 132505
Chronology: Jun. 2024
DOI: 10.1016/j.ijbiomac.2024.132505
ID: 24230211