| Autor: |
Monte JFS; Programa de Pós-graduação em Bioquímica, Centro de Biociências, Universidade Federal do Rio Grande do Norte, Natal, Brazil., Moreno CJG; Programa de Pós-graduação em Bioquímica, Centro de Biociências, Universidade Federal do Rio Grande do Norte, Natal, Brazil., Monteiro JPMFL; Global Health and Tropical Medicine, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisbon, Portugal., de Oliveira Rocha HA; Programa de Pós-graduação em Bioquímica, Centro de Biociências, Universidade Federal do Rio Grande do Norte, Natal, Brazil., Ribeiro AR; Departmento de Parasitologia, Universidade Estadual de Campinas, Campinas, 13083-862, Brazil., Silva MS; Programa de Pós-graduação em Bioquímica, Centro de Biociências, Universidade Federal do Rio Grande do Norte, Natal, Brazil. mssilva.ufrn@gmail.com.; Global Health and Tropical Medicine, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisbon, Portugal. mssilva.ufrn@gmail.com.; Departamento de Análises Clínicas e Toxicológicas, Centro de Ciências da Saúde, Universidade Federal do Rio Grande do Norte, Natal, Brazil. mssilva.ufrn@gmail.com. |
| Abstrakt: |
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. |
