Abstract
Zymography assay is a semiquantitative technique, very sensitive, and commonly used to determine metalloproteinase levels in different types of biological samples, including tissues, cells, and extracts of protein. Samples containing metalloproteinases are loaded onto a polyacrylamide gel containing sodium dodecyl sulphate (SDS) and a specific substrate (gelatin, casein, collagen, etc.). Then proteins are allowed to migrate under an electric current and the distance of migration is inversely correlated with the molecular weight. After migration, the gel is placed in a renaturing buffer to allow proteins to regain their tertiary structure, necessary for enzymatic activity (metalloproteinase activity). In the context of infections caused by trypanosomatids (Leishmania spp., Trypanosoma cruzi, and Trypanosoma brucei), the characterization of metalloproteinase by zymography can contribute to the comprehension of the pathogenesis mechanisms and host–parasite interaction.
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Acknowledgments
This work was supported by Capes—Brazil (Bolsa Jovem Talento—Grant 019/2013) and by Global Health and Tropical Medicine (GHTM-UID/multi/04413/2013). We wish to thank Dr. João Aristeu da Rosa from Faculdade de Ciências Farmacêuticas de Araraquara—Brazil, who isolated the QMM5 strain of Trypanosoma cruzi parasite.
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Monte, J.F.S., Moreno, C.J.G., Monteiro, J.P.M.F.L., de Oliveira Rocha, H.A., Ribeiro, A.R., Silva, M.S. (2017). Use of Zymography in Trypanosomiasis Studies. In: Wilkesman, J., Kurz, L. (eds) Zymography. Methods in Molecular Biology, vol 1626. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7111-4_20
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DOI: https://doi.org/10.1007/978-1-4939-7111-4_20
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