Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis
With the aim to study proteinases released to the culture medium during Trypanosoma cruzi metacyclogenesis, the presence of cysteine proteinases (CPs) was analysed in culture supernatants obtained throughout the differentiation induced by stimulation of epimastigotes with Triatoma infestans hindgut...
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todo:paper_00311820_v132_n3_p345_Duschak2023-10-03T14:41:13Z Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis Duschak, V.G. Barboza, M. García, G.A. Lammel, E.M. Couto, A.S. Isola, E.L.D. Cruzipain Cysteine proteinases Metacyclogenesis Trypanosoma cruzi concanavalin A cruzipain cysteine proteinase n [n (3 carboxyoxirane 2 carbonyl)leucyl]agmatine tosyllysyl chloromethyl ketone animal tissue article binding affinity controlled study differentiation enzyme activity enzyme isolation enzyme substrate complex epimastigote growth curve life cycle nonhuman priority journal Triatoma infestans trypomastigote Animals Blotting, Western Chromatography, Affinity Cross Reactions Culture Media Culture Techniques Cysteine Endopeptidases Hydrogen-Ion Concentration Life Cycle Stages Protease Inhibitors Triatoma Trypanosoma cruzi Animalia Triatoma infestans Trypanosoma cruzi With the aim to study proteinases released to the culture medium during Trypanosoma cruzi metacyclogenesis, the presence of cysteine proteinases (CPs) was analysed in culture supernatants obtained throughout the differentiation induced by stimulation of epimastigotes with Triatoma infestans hindgut homogenate. In SDS-gelatin containing gels, an important endopeptidase activity with apparent molecular weight range between 97 and 116 kDa was encountered at pH 6, which was abolished by the specific cysteine proteinase inhibitor E-64 and TLCK, but not by pepstatin, 1,10 phenantroline or PMSF. This novel CP, named TcCPmet, showed affinity to cystatin-Sepharose, denoting its thiol-proteinase character as well as to ConA-Sepharose, indicating it contains N-linked oligosaccharides. However, it presented a different elution pattern on ConA-Sepharose than cruzipain and, in addition, it was not recognized by anti-cruzipain serum, facts that strongly suggest the different nature of both CPs. Moroever, evidence is presented indicating that TcCPmet was able to hydrolyse the same chromogenic peptides as cruzipain at optimal alkaline pH values, although with a different order of effectiveness. Our results indicate the presence of a novel CP secreted by metacyclic trypomastigotes and reinforces the important role of these enzymes in metacyclogenesis. © 2005 Cambridge University Press. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00311820_v132_n3_p345_Duschak |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
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R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Cruzipain Cysteine proteinases Metacyclogenesis Trypanosoma cruzi concanavalin A cruzipain cysteine proteinase n [n (3 carboxyoxirane 2 carbonyl)leucyl]agmatine tosyllysyl chloromethyl ketone animal tissue article binding affinity controlled study differentiation enzyme activity enzyme isolation enzyme substrate complex epimastigote growth curve life cycle nonhuman priority journal Triatoma infestans trypomastigote Animals Blotting, Western Chromatography, Affinity Cross Reactions Culture Media Culture Techniques Cysteine Endopeptidases Hydrogen-Ion Concentration Life Cycle Stages Protease Inhibitors Triatoma Trypanosoma cruzi Animalia Triatoma infestans Trypanosoma cruzi |
| spellingShingle |
Cruzipain Cysteine proteinases Metacyclogenesis Trypanosoma cruzi concanavalin A cruzipain cysteine proteinase n [n (3 carboxyoxirane 2 carbonyl)leucyl]agmatine tosyllysyl chloromethyl ketone animal tissue article binding affinity controlled study differentiation enzyme activity enzyme isolation enzyme substrate complex epimastigote growth curve life cycle nonhuman priority journal Triatoma infestans trypomastigote Animals Blotting, Western Chromatography, Affinity Cross Reactions Culture Media Culture Techniques Cysteine Endopeptidases Hydrogen-Ion Concentration Life Cycle Stages Protease Inhibitors Triatoma Trypanosoma cruzi Animalia Triatoma infestans Trypanosoma cruzi Duschak, V.G. Barboza, M. García, G.A. Lammel, E.M. Couto, A.S. Isola, E.L.D. Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis |
| topic_facet |
Cruzipain Cysteine proteinases Metacyclogenesis Trypanosoma cruzi concanavalin A cruzipain cysteine proteinase n [n (3 carboxyoxirane 2 carbonyl)leucyl]agmatine tosyllysyl chloromethyl ketone animal tissue article binding affinity controlled study differentiation enzyme activity enzyme isolation enzyme substrate complex epimastigote growth curve life cycle nonhuman priority journal Triatoma infestans trypomastigote Animals Blotting, Western Chromatography, Affinity Cross Reactions Culture Media Culture Techniques Cysteine Endopeptidases Hydrogen-Ion Concentration Life Cycle Stages Protease Inhibitors Triatoma Trypanosoma cruzi Animalia Triatoma infestans Trypanosoma cruzi |
| description |
With the aim to study proteinases released to the culture medium during Trypanosoma cruzi metacyclogenesis, the presence of cysteine proteinases (CPs) was analysed in culture supernatants obtained throughout the differentiation induced by stimulation of epimastigotes with Triatoma infestans hindgut homogenate. In SDS-gelatin containing gels, an important endopeptidase activity with apparent molecular weight range between 97 and 116 kDa was encountered at pH 6, which was abolished by the specific cysteine proteinase inhibitor E-64 and TLCK, but not by pepstatin, 1,10 phenantroline or PMSF. This novel CP, named TcCPmet, showed affinity to cystatin-Sepharose, denoting its thiol-proteinase character as well as to ConA-Sepharose, indicating it contains N-linked oligosaccharides. However, it presented a different elution pattern on ConA-Sepharose than cruzipain and, in addition, it was not recognized by anti-cruzipain serum, facts that strongly suggest the different nature of both CPs. Moroever, evidence is presented indicating that TcCPmet was able to hydrolyse the same chromogenic peptides as cruzipain at optimal alkaline pH values, although with a different order of effectiveness. Our results indicate the presence of a novel CP secreted by metacyclic trypomastigotes and reinforces the important role of these enzymes in metacyclogenesis. © 2005 Cambridge University Press. |
| format |
JOUR |
| author |
Duschak, V.G. Barboza, M. García, G.A. Lammel, E.M. Couto, A.S. Isola, E.L.D. |
| author_facet |
Duschak, V.G. Barboza, M. García, G.A. Lammel, E.M. Couto, A.S. Isola, E.L.D. |
| author_sort |
Duschak, V.G. |
| title |
Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis |
| title_short |
Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis |
| title_full |
Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis |
| title_fullStr |
Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis |
| title_full_unstemmed |
Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis |
| title_sort |
novel cysteine proteinase in trypanosoma cruzi metacyclogenesis |
| url |
http://hdl.handle.net/20.500.12110/paper_00311820_v132_n3_p345_Duschak |
| work_keys_str_mv |
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| _version_ |
1782024487532756992 |