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Expression and Purification of Active Monomeric MMP7
Kazuhiro Yamamoto1, Moe Isohata2, Shouichi Higashi3
1Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool, UK.
Abstract:
MMP7 is the smallest member of the MMP family and plays multiple physiological and pathological roles through interaction with a variety of molecules. Purified MMP7 would be beneficial for studying its function and for the development of inhibitors, which could be potential therapeutics. Due to low levels of endogenously produced MMP7, its recombinant expression and purification using E. coli have been established. Here, we describe an effective method to express and purify an active form of MMP7. Our recent discovery is that adding high concentration of CaCl2 during refolding process prevents nonspecific binding of MMP7 to plastic and its aggregation, significantly improving the yield of active monomeric forms of MMP7.
Insights
Researchers developed a new method to produce active Matrix Metalloproteinase 7 (MMP7). Adding calcium chloride during refolding significantly boosted the yield of pure, active MMP7 for therapeutic research.
Area of Science:
- Biochemistry
- Molecular Biology
- Protein Chemistry
Background:
- Matrix Metalloproteinase 7 (MMP7) is the smallest MMP, involved in various physiological and pathological processes.
- Recombinant MMP7 production is crucial for functional studies and therapeutic inhibitor development.
- Existing recombinant MMP7 expression methods face challenges with low yields and purity.
Purpose of the Study:
- To establish an effective method for expressing and purifying an active form of MMP7.
- To improve the yield and purity of active, monomeric MMP7 for research applications.
Main Methods:
- Recombinant expression of MMP7 in E. coli.
- Optimization of the refolding process using high concentrations of CaCl2.
- Purification of active monomeric MMP7.
Main Results:
- A novel refolding strategy using high CaCl2 concentration was identified.
- This method effectively prevented non-specific binding and aggregation of MMP7.
- Significantly improved yields of active, monomeric MMP7 were achieved.
Conclusions:
- The described method provides an efficient way to obtain active MMP7.
- This advancement facilitates further research into MMP7's roles and the development of MMP7-targeted therapeutics.
