Targeted approach to determine the impact of cancer-associated protease variants
Several steps of cancer progression, from tumor onset to metastasis, critically involve proteolytic activity. To elucidate the role of proteases in cancer, it is particularly important to consider single-nucleotide variants (SNVs) that affect the active site of proteases, thereby influencing cleavage specificity, substrate processing, and thus cancer cell behavior. To facilitate systematic studies, we here present a targeted approach to determine the impact of cancer-associated protease variants (TACAP). Starting with the semiautomated identification of potential specificity-modulating SNVs, our workflow comprises mass spectrometry-based cleavage specificity profiling and substrate identification, localization, and inhibitor studies, followed by functional analyses investigating cancer cell properties. To demonstrate the feasibility of TACAP, we analyzed the meprin β R238Q variant. This amino acid exchange R238Q leads to a loss of meprin β's characteristic cleavage preference for acidic amino acids at P1' position, accompanied with changes in substrate pool and inhibitor affinity compared to meprin β wild type.
Authors
- Claus U. Pietrzik (ORCID: https://orcid.org/0000-0002-1202-5489)
- Franka Scharfenberg
- Dimitris Georgiadis (ORCID: https://orcid.org/0000-0002-9656-0701)
- Fred Armbrust (ORCID: https://orcid.org/0000-0002-3331-5522)
- Tomas Koudelka
- Sascha Rahn
- Kira Bickenbach (ORCID: https://orcid.org/0000-0001-5079-0512)
- Fabrice Beau
- Andreas Tholey (ORCID: https://orcid.org/0000-0002-8687-6817)
- Noel David (ORCID: https://orcid.org/0009-0009-3589-8264)
- Christoph Becker‐Pauly (ORCID: https://orcid.org/0000-0002-5100-9916)
- Lea Stahmer
- Malina Rüffer
- Corentin Joos
Institutions
- Johannes Gutenberg University Mainz (DE)
- National and Kapodistrian University of Athens (GR)
- Commissariat à l'Énergie Atomique et aux Énergies Alternatives (FR)
- Université Paris-Saclay (FR)
- Christian-Albrechts-Universität zu Kiel (DE)
- Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement (FR)
- University Medical Center of the Johannes Gutenberg University Mainz (DE)
- Technologies pour la Santé (FR)
- CEA Paris-Saclay (FR)
Publication Details
- Journal
- Science Advances
- Published
- 2025-02-12
- DOI
- https://doi.org/10.1126/sciadv.adp5958
- Citations
- 2
- Primary Topic
- Advanced Proteomics Techniques and Applications
- Type
- article
- Field-Weighted Citation Impact
- 1.51
Funders
- Universitätsmedizin Göttingen