With the recent release of the first commercially available and ready-to-use assay to detect MMP-25 activity , Enzium has opened a new era for screening protease activities through a robust non-FRET experimental approach. A good opportunity to take a look at the EnSens technology.
Unlike other members of the family of Matrix MetalloProteases (MMPs) which are secreted as inactive pro-proteins and activated upon cleavage of extracellular enzymes, MMP-25 (or MT-MMP 6; formerly called MMP-20) (1) is attached to the plasma membrane. It is thought that MMP-25 inactivates alpha-1 Proteinase Inhibitor which serves as a tissue protectant against proteolytic enzymes released by activated neutrophils in response to bacterial infection or inflammation. Thus the protein might play a role in facilitating transendothelial migration of neutrophils to inflammatory sites. Furthermore, MMP-25 might play a role in tumor invasion and metastasis through activation on MMP-2 (2).
Hence MMP-25 should be seen as a potential target in Drug Discovery. Until now no activity assay has been available for MMP-25.
MMP-25 ready-to-use activity assay kit
Recently, Enzium release the first ready-to-use assay kit for measuring MMP-25 activity.
Several lines of evidence prove that this kit is an ideal companion for measuring MMP-25 activity and for screening inhibitors.
The EnSens substrate can incorporate full recognition site sequences – thus the selectivity of the assays is superior as compared to classical FRET-based technology. The substrate used is modular, allowing easy swap of protease recognition sites. Interestingly, customized substrates can even be designed if you know the recognition site of your protease of interest!
The EnSens se-Red fluorophore is resistant to photobleaching up to 48 hours. The long emission wavelength (665nm) generated reduces interference from other assay components ensuring optimal signal-to-noise ratios (6-20x).
Enzium’s scientists have proven that their assay is highly selective for the biological mimicking MMP-25 and MMP-17 (both belonging to the MT-MMP subfamily). To do so, they challenged their MMP-25 activity assay with substrates related to various proteases such as MMP-9 and MMP-8.
Interested in measuring MMP activity?
Various proteases and MMPs are now eligible for activity measurement and inhibitor screening via the EnSens technology. You can thus benefit from this unique from this technology in your lab and even subcontract your EnSens Protease activity assays and inhibitor screening to external laboratories such as tebu-bio laboratories.
Looking for a protease with a known recognition site, planning a new screening program in-house or via outsourcing? Leave a message in the form below.
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Sources:
- Pei D., Cell Res 9 (4): 291–303 (2000)
- Entrez Gene: MMP25 matrix metallopeptidase 25
