Analysis of Peptidases in Non-Infected and Trypanosoma cruzi-Infected Mouse Embryo Hepatocyte Cells
Cellular and extracellular peptidase profiles from non-infected and Trypanosoma cruzi-infected hepatocyte cell cultures were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) containing different copolymerized proteins as substrates. A 100 kDa metallopeptidase act...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Master Publishing Group
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3614691/ https://www.ncbi.nlm.nih.gov/pubmed/23675074 |
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author | de Melo, Ana Cristina Nogueira dos Santos, André Luis Souza Leal Meirelles, Maria Nazareth Branquinha, Marta Helena Vermelho, Alane Beatriz |
author_facet | de Melo, Ana Cristina Nogueira dos Santos, André Luis Souza Leal Meirelles, Maria Nazareth Branquinha, Marta Helena Vermelho, Alane Beatriz |
author_sort | de Melo, Ana Cristina Nogueira |
collection | PubMed |
description | Cellular and extracellular peptidase profiles from non-infected and Trypanosoma cruzi-infected hepatocyte cell cultures were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) containing different copolymerized proteins as substrates. A 100 kDa metallopeptidase activity was detected in the cellular extracts and in the culture supernatant fluids of both systems, had the ability to exclusively degrade gelatin. However, non-infected hepatocytes produced an additional extracellular metallopeptidase of 85 kDa. In the non-infected and in the infected hepatocytes, a cysteine peptidase migrating in gelatin-SDS-PAGE at 60 kDa presented the broadest specificity, since it was also able to hydrolyze casein and hemoglobin. The 100 kDa component was only detected at alkaline pH and predominantly expressed in non-infected hepatocytes. Conversely, the 60 kDa cysteine peptidase was only observed in acidic condition and its production was robustly augmented in T. cruzi-infected cells, probably due to the cysteine peptidase synthesized by the parasites, as corroborated by immunoblotting assay using anti-cruzipain antibody. Collectively, these results suggest that peptidases may be involved in the interaction process between T. cruzi and hepatocytes in vitro. |
format | Online Article Text |
id | pubmed-3614691 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | Master Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-36146912013-05-01 Analysis of Peptidases in Non-Infected and Trypanosoma cruzi-Infected Mouse Embryo Hepatocyte Cells de Melo, Ana Cristina Nogueira dos Santos, André Luis Souza Leal Meirelles, Maria Nazareth Branquinha, Marta Helena Vermelho, Alane Beatriz Int J Biomed Sci Article Cellular and extracellular peptidase profiles from non-infected and Trypanosoma cruzi-infected hepatocyte cell cultures were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) containing different copolymerized proteins as substrates. A 100 kDa metallopeptidase activity was detected in the cellular extracts and in the culture supernatant fluids of both systems, had the ability to exclusively degrade gelatin. However, non-infected hepatocytes produced an additional extracellular metallopeptidase of 85 kDa. In the non-infected and in the infected hepatocytes, a cysteine peptidase migrating in gelatin-SDS-PAGE at 60 kDa presented the broadest specificity, since it was also able to hydrolyze casein and hemoglobin. The 100 kDa component was only detected at alkaline pH and predominantly expressed in non-infected hepatocytes. Conversely, the 60 kDa cysteine peptidase was only observed in acidic condition and its production was robustly augmented in T. cruzi-infected cells, probably due to the cysteine peptidase synthesized by the parasites, as corroborated by immunoblotting assay using anti-cruzipain antibody. Collectively, these results suggest that peptidases may be involved in the interaction process between T. cruzi and hepatocytes in vitro. Master Publishing Group 2008-06 /pmc/articles/PMC3614691/ /pubmed/23675074 Text en © Ana Cristina Melo et al. Licensee Master Publishing Group http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.5/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Article de Melo, Ana Cristina Nogueira dos Santos, André Luis Souza Leal Meirelles, Maria Nazareth Branquinha, Marta Helena Vermelho, Alane Beatriz Analysis of Peptidases in Non-Infected and Trypanosoma cruzi-Infected Mouse Embryo Hepatocyte Cells |
title | Analysis of Peptidases in Non-Infected and Trypanosoma cruzi-Infected Mouse Embryo Hepatocyte Cells |
title_full | Analysis of Peptidases in Non-Infected and Trypanosoma cruzi-Infected Mouse Embryo Hepatocyte Cells |
title_fullStr | Analysis of Peptidases in Non-Infected and Trypanosoma cruzi-Infected Mouse Embryo Hepatocyte Cells |
title_full_unstemmed | Analysis of Peptidases in Non-Infected and Trypanosoma cruzi-Infected Mouse Embryo Hepatocyte Cells |
title_short | Analysis of Peptidases in Non-Infected and Trypanosoma cruzi-Infected Mouse Embryo Hepatocyte Cells |
title_sort | analysis of peptidases in non-infected and trypanosoma cruzi-infected mouse embryo hepatocyte cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3614691/ https://www.ncbi.nlm.nih.gov/pubmed/23675074 |
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