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.
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Acknowledgments
This work was supported in part by an Extramural Collaborative Research Grant of the Cancer Research Institute, Kanazawa University, Japan (to S. H.), and Grants-in-Aid for Scientific Research (C) Grant Number JP19K07047 (to S. H.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan. K. Y. was supported by the Versus Arthritis (21447 and 23137).
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Yamamoto, K., Isohata, M., Higashi, S. (2024). Expression and Purification of Active Monomeric MMP7. In: Santamaria, S. (eds) Proteases and Cancer. Methods in Molecular Biology, vol 2747. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3589-6_6
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DOI: https://doi.org/10.1007/978-1-0716-3589-6_6
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