TY  - JOUR
A1  - Wagner, Niklas
A1  - Rapp, Anna Elise
A1  - Braun, Sebastian Benedict
A1  - Ehnert, Markus
A1  - Imhof, Thomas
A1  - Koch, Manuel
A1  - Jenei-Lanzl, Zsuzsa
A1  - Zaucke, Frank
A1  - Meurer, Andrea
T1  - Generation of matrix degradation products using an in vitro MMP cleavage assay
T2  - International journal of molecular sciences
N2  - 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.
KW  - MMPs
KW  - cartilage
KW  - COMP
KW  - thrombospondins
KW  - osteoarthritis
Y1  - 2022
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/69248
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-692484
SN  - 1422-0067
N1  - 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)).
VL  - 23
IS  - 11, art. 6245
SP  - 1
EP  - 18
PB  - Molecular Diversity Preservation International
CY  - Basel
ER  -