Effects of chlorate on the sulfation process of trypanosoma cruzi glycoconjugates. Implication of parasite sulfates in cellular invasion
Sulfation, a post-translational modification which plays a key role in various biological processes, is inhibited by competition with chlorate. In Trypanosoma cruzi, the agent of Chagas' disease, sulfated structures have been described as part of glycolipids and we have reported sulfated high-m...
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todo:paper_0001706X_v137_n_p161_Ferrero2023-10-03T13:51:39Z Effects of chlorate on the sulfation process of trypanosoma cruzi glycoconjugates. Implication of parasite sulfates in cellular invasion Ferrero, M.R. Soprano, L.L. Acosta, D.M. García, G.A. Esteva, M.I. Couto, A.S. Duschak, V.G. Cruzipain Invasion process Sulfation Sulfoglycosphingolipids Trypanosoma cruzi chlorate cruzipain cruzipain antibody enzyme antibody glycoconjugate immunoglobulin G mannose receptor mannose receptor antibody receptor antibody sulfatide unclassified drug chlorate cysteine proteinase glycoconjugate sulfate antibody electron microscopy lipid membrane metabolism parasite amastigote animal cell antibody specificity article cell invasion cell organelle cell ultrastructure controlled study electron microscopy epimastigote flow cytometry gel mobility shift assay host parasite interaction immunoblotting isoelectric point lipid reservosome matrix assisted laser desorption ionization time of flight mass spectrometry nonhuman polyacrylamide gel electrophoresis sulfation thin layer chromatography Trypanosoma cruzi trypomastigote ultraviolet spectroscopy animal cell line drug effects endocytosis heart muscle cell human mass spectrometry metabolism parasitology physiology protein processing rabbit Trypanosoma cruzi two dimensional gel electrophoresis Animals Cell Line Chlorates Cysteine Endopeptidases Electrophoresis, Gel, Two-Dimensional Endocytosis Glycoconjugates Humans Immunoblotting Isoelectric Point Metabolic Networks and Pathways Microscopy, Electron Myocytes, Cardiac Protein Processing, Post-Translational Rabbits Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Sulfates Trypanosoma cruzi Sulfation, a post-translational modification which plays a key role in various biological processes, is inhibited by competition with chlorate. In Trypanosoma cruzi, the agent of Chagas' disease, sulfated structures have been described as part of glycolipids and we have reported sulfated high-mannose type oligosaccharides in the C-T domain of the cruzipain (Cz) glycoprotein. However, sulfation pathways have not been described yet in this parasite. Herein, we studied the effect of chlorate treatment on T. cruzi with the aim to gain some knowledge about sulfation metabolism and the role of sulfated molecules in this parasite. In chlorate-treated epimastigotes, immunoblotting with anti-sulfates enriched Cz IgGs (AS-enriched IgGs) showed Cz undersulfation. Accordingly, a Cz mobility shift toward higher isoelectric points was observed in 2D-PAGE probed with anti-Cz antibodies. Ultrastructural membrane abnormalities and a significant decrease of dark lipid reservosomes were shown by electron microscopy and a significant decrease in sulfatide levels was confirmed by TLC/UV-MALDI-TOF-MS analysis. Altogether, these results suggest T. cruzi sulfation occurs via PAPS. Sulfated epitopes in trypomastigote and amastigote forms were evidenced using AS-enriched IgGs by immunoblotting. Their presence on trypomastigotes surface was demonstrated by flow cytometry and IF with Cz/dCz specific antibodies. Interestingly, the percentage of infected cardiac HL-1 cells decreased 40% when using chlorate-treated trypomastigotes, suggesting sulfates are involved in the invasion process. The same effect was observed when cells were pre-incubated with dCz, dC-T or an anti-high mannose receptor (HMR) antibody, suggesting Cz sulfates and HMR are also involved in the infection process by T. cruzi. © 2014 Elsevier B.V. Fil:García, G.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Couto, A.S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_0001706X_v137_n_p161_Ferrero |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Cruzipain Invasion process Sulfation Sulfoglycosphingolipids Trypanosoma cruzi chlorate cruzipain cruzipain antibody enzyme antibody glycoconjugate immunoglobulin G mannose receptor mannose receptor antibody receptor antibody sulfatide unclassified drug chlorate cysteine proteinase glycoconjugate sulfate antibody electron microscopy lipid membrane metabolism parasite amastigote animal cell antibody specificity article cell invasion cell organelle cell ultrastructure controlled study electron microscopy epimastigote flow cytometry gel mobility shift assay host parasite interaction immunoblotting isoelectric point lipid reservosome matrix assisted laser desorption ionization time of flight mass spectrometry nonhuman polyacrylamide gel electrophoresis sulfation thin layer chromatography Trypanosoma cruzi trypomastigote ultraviolet spectroscopy animal cell line drug effects endocytosis heart muscle cell human mass spectrometry metabolism parasitology physiology protein processing rabbit Trypanosoma cruzi two dimensional gel electrophoresis Animals Cell Line Chlorates Cysteine Endopeptidases Electrophoresis, Gel, Two-Dimensional Endocytosis Glycoconjugates Humans Immunoblotting Isoelectric Point Metabolic Networks and Pathways Microscopy, Electron Myocytes, Cardiac Protein Processing, Post-Translational Rabbits Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Sulfates Trypanosoma cruzi |
| spellingShingle |
Cruzipain Invasion process Sulfation Sulfoglycosphingolipids Trypanosoma cruzi chlorate cruzipain cruzipain antibody enzyme antibody glycoconjugate immunoglobulin G mannose receptor mannose receptor antibody receptor antibody sulfatide unclassified drug chlorate cysteine proteinase glycoconjugate sulfate antibody electron microscopy lipid membrane metabolism parasite amastigote animal cell antibody specificity article cell invasion cell organelle cell ultrastructure controlled study electron microscopy epimastigote flow cytometry gel mobility shift assay host parasite interaction immunoblotting isoelectric point lipid reservosome matrix assisted laser desorption ionization time of flight mass spectrometry nonhuman polyacrylamide gel electrophoresis sulfation thin layer chromatography Trypanosoma cruzi trypomastigote ultraviolet spectroscopy animal cell line drug effects endocytosis heart muscle cell human mass spectrometry metabolism parasitology physiology protein processing rabbit Trypanosoma cruzi two dimensional gel electrophoresis Animals Cell Line Chlorates Cysteine Endopeptidases Electrophoresis, Gel, Two-Dimensional Endocytosis Glycoconjugates Humans Immunoblotting Isoelectric Point Metabolic Networks and Pathways Microscopy, Electron Myocytes, Cardiac Protein Processing, Post-Translational Rabbits Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Sulfates Trypanosoma cruzi Ferrero, M.R. Soprano, L.L. Acosta, D.M. García, G.A. Esteva, M.I. Couto, A.S. Duschak, V.G. Effects of chlorate on the sulfation process of trypanosoma cruzi glycoconjugates. Implication of parasite sulfates in cellular invasion |
| topic_facet |
Cruzipain Invasion process Sulfation Sulfoglycosphingolipids Trypanosoma cruzi chlorate cruzipain cruzipain antibody enzyme antibody glycoconjugate immunoglobulin G mannose receptor mannose receptor antibody receptor antibody sulfatide unclassified drug chlorate cysteine proteinase glycoconjugate sulfate antibody electron microscopy lipid membrane metabolism parasite amastigote animal cell antibody specificity article cell invasion cell organelle cell ultrastructure controlled study electron microscopy epimastigote flow cytometry gel mobility shift assay host parasite interaction immunoblotting isoelectric point lipid reservosome matrix assisted laser desorption ionization time of flight mass spectrometry nonhuman polyacrylamide gel electrophoresis sulfation thin layer chromatography Trypanosoma cruzi trypomastigote ultraviolet spectroscopy animal cell line drug effects endocytosis heart muscle cell human mass spectrometry metabolism parasitology physiology protein processing rabbit Trypanosoma cruzi two dimensional gel electrophoresis Animals Cell Line Chlorates Cysteine Endopeptidases Electrophoresis, Gel, Two-Dimensional Endocytosis Glycoconjugates Humans Immunoblotting Isoelectric Point Metabolic Networks and Pathways Microscopy, Electron Myocytes, Cardiac Protein Processing, Post-Translational Rabbits Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Sulfates Trypanosoma cruzi |
| description |
Sulfation, a post-translational modification which plays a key role in various biological processes, is inhibited by competition with chlorate. In Trypanosoma cruzi, the agent of Chagas' disease, sulfated structures have been described as part of glycolipids and we have reported sulfated high-mannose type oligosaccharides in the C-T domain of the cruzipain (Cz) glycoprotein. However, sulfation pathways have not been described yet in this parasite. Herein, we studied the effect of chlorate treatment on T. cruzi with the aim to gain some knowledge about sulfation metabolism and the role of sulfated molecules in this parasite. In chlorate-treated epimastigotes, immunoblotting with anti-sulfates enriched Cz IgGs (AS-enriched IgGs) showed Cz undersulfation. Accordingly, a Cz mobility shift toward higher isoelectric points was observed in 2D-PAGE probed with anti-Cz antibodies. Ultrastructural membrane abnormalities and a significant decrease of dark lipid reservosomes were shown by electron microscopy and a significant decrease in sulfatide levels was confirmed by TLC/UV-MALDI-TOF-MS analysis. Altogether, these results suggest T. cruzi sulfation occurs via PAPS. Sulfated epitopes in trypomastigote and amastigote forms were evidenced using AS-enriched IgGs by immunoblotting. Their presence on trypomastigotes surface was demonstrated by flow cytometry and IF with Cz/dCz specific antibodies. Interestingly, the percentage of infected cardiac HL-1 cells decreased 40% when using chlorate-treated trypomastigotes, suggesting sulfates are involved in the invasion process. The same effect was observed when cells were pre-incubated with dCz, dC-T or an anti-high mannose receptor (HMR) antibody, suggesting Cz sulfates and HMR are also involved in the infection process by T. cruzi. © 2014 Elsevier B.V. |
| format |
JOUR |
| author |
Ferrero, M.R. Soprano, L.L. Acosta, D.M. García, G.A. Esteva, M.I. Couto, A.S. Duschak, V.G. |
| author_facet |
Ferrero, M.R. Soprano, L.L. Acosta, D.M. García, G.A. Esteva, M.I. Couto, A.S. Duschak, V.G. |
| author_sort |
Ferrero, M.R. |
| title |
Effects of chlorate on the sulfation process of trypanosoma cruzi glycoconjugates. Implication of parasite sulfates in cellular invasion |
| title_short |
Effects of chlorate on the sulfation process of trypanosoma cruzi glycoconjugates. Implication of parasite sulfates in cellular invasion |
| title_full |
Effects of chlorate on the sulfation process of trypanosoma cruzi glycoconjugates. Implication of parasite sulfates in cellular invasion |
| title_fullStr |
Effects of chlorate on the sulfation process of trypanosoma cruzi glycoconjugates. Implication of parasite sulfates in cellular invasion |
| title_full_unstemmed |
Effects of chlorate on the sulfation process of trypanosoma cruzi glycoconjugates. Implication of parasite sulfates in cellular invasion |
| title_sort |
effects of chlorate on the sulfation process of trypanosoma cruzi glycoconjugates. implication of parasite sulfates in cellular invasion |
| url |
http://hdl.handle.net/20.500.12110/paper_0001706X_v137_n_p161_Ferrero |
| work_keys_str_mv |
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