Generation of matrix degradation products using an in vitro MMP cleavage assay
- Matrix metalloproteinases (MMPs) play crucial roles in tissue homeostasis and pathologies by remodeling the extracellular matrix. Previous studies have demonstrated the biological activities of MMP-derived cleavage products. Furthermore, specific fragments can serve as biomarkers. Therefore, an in vitro cleavage assay to identify substrates and characterize cleavage patterns could provide important insight in disease-relevant mechanisms and the identification of novel biomarkers. In the pathogenesis of osteoarthritis (OA), MMP-2, -8, -9 and -13 are of vital importance. However, it is unclear which protease can cleave which matrix component. To address this question, we established an in vitro cleavage assay using recombinantly expressed MMPs and the two cartilage matrix components, COMP and thrombospondin-4. We found a time- and concentration-dependent degradation and an MMP-specific cleavage pattern for both proteins. Cleavage products can now be enriched and purified to investigate their biological activity. To verify the in vivo relevance, we compared the in vitro cleavage patterns with serum and synovial fluid from OA patients and could indeed detect fragments of similar size in the human samples. The cleavage assay can be adapted to other MMPs and substrates, making it a valuable tool for many research fields.
Author: | Niklas Wagner, Anna Elise RappORCiDGND, Sebastian Benedict BraunORCiDGND, Markus EhnertGND, Thomas ImhofGND, Manuel KochORCiDGND, Zsuzsa Jenei-LanzlORCiDGND, Frank ZauckeORCiDGND, Andrea MeurerORCiD |
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URN: | urn:nbn:de:hebis:30:3-692484 |
DOI: | https://doi.org/10.3390/ijms23116245 |
ISSN: | 1422-0067 |
Parent Title (English): | International journal of molecular sciences |
Publisher: | Molecular Diversity Preservation International |
Place of publication: | Basel |
Document Type: | Article |
Language: | English |
Date of Publication (online): | 2022/06/02 |
Date of first Publication: | 2022/06/02 |
Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
Release Date: | 2022/12/12 |
Tag: | COMP; MMPs; cartilage; osteoarthritis; thrombospondins |
Volume: | 23 |
Issue: | 11, art. 6245 |
Article Number: | 6245 |
Page Number: | 18 |
First Page: | 1 |
Last Page: | 18 |
Note: | This research was funded by the Deutsche Forschungsgemeinschaft FOR2407 (JE 642/4-2 to Z.J.-L. (project number 277277765)), and FOR2722 (FZ 561/3-1 to F.Z. (project number 407168728) and to M.K. (project number 384170921)). |
HeBIS-PPN: | 505601583 |
Institutes: | Medizin |
Dewey Decimal Classification: | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
Sammlungen: | Universitätspublikationen |
Licence (German): | ![]() |