Use of Zymography in Trypanosomiasis Studies

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1626; p. 213
• Main Authors: Monte, Jéssyka Fernanda Santiago, Moreno, Cláudia Jassica Gonçalves, Monteiro, Joana Patrícia Molato Figueiredo Lopes, de Oliveira Rocha, Hugo Alexandre, Ribeiro, Aline Rimoldi, Silva, Marcelo Sousa
• Format: Journal Article
• Language: English
• Published: United States 2017
• Subjects: Animals; Collagen - metabolism; Electrophoresis, Polyacrylamide Gel - methods; Enzyme Assays - methods; Gelatin - metabolism; Host-Parasite Interactions; Humans; Leishmania - enzymology; Leishmania - metabolism; Leishmaniasis - metabolism; Leishmaniasis - parasitology; Metalloendopeptidases - analysis; Metalloendopeptidases - metabolism; Trypanosoma brucei brucei - enzymology; Trypanosoma brucei brucei - metabolism; Trypanosoma cruzi - enzymology; Trypanosoma cruzi - metabolism; Trypanosomiasis - metabolism; Trypanosomiasis - parasitology; Zymography; Trypanosomatids; Metalloproteinases; Trypanosoma brucei; Leishmania spp; Trypanosoma cruzi
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-7111-4_20

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.