Cyclic AMP-dependent protein kinase activity in Trypanosoma cruzi

A cyclic AMP-dependent protein kinase activity from epimastigote forms of Trypanosoma cruzi was characterized. Cytosolic extracts were chromatographed on DEAE-cellulose columns, giving two peaks of kinase activity, which were eluted at 0.15 M- and 0.32 M-NaCl respectively. The second activity peak w...

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Autores principales: Ulloa, R.M., Mesri, E., Esteva, M., Torres, H.N., Tellez-Inon, M.T.
Formato: JOUR
Materias:
antibody
cyclic amp
protein kinase
immunoblotting
nonhuman
priority journal
protozoon
trypanosoma cruzi
Animal
Cattle
Chromatography, Affinity
Chromatography, DEAE-Cellulose
Chromatography, Gel
Cyclic AMP
Immunoblotting
Myocardium
Phosphates
Phosphorylation
Protein Binding
Protein Kinases
Support, Non-U.S. Gov't
Trypanosoma cruzi
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_02646021_v255_n1_p319_Ulloa
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
id todo:paper_02646021_v255_n1_p319_Ulloa
record_format dspace
spelling todo:paper_02646021_v255_n1_p319_Ulloa2023-10-03T15:12:55Z Cyclic AMP-dependent protein kinase activity in Trypanosoma cruzi Ulloa, R.M. Mesri, E. Esteva, M. Torres, H.N. Tellez-Inon, M.T. antibody cyclic amp protein kinase immunoblotting nonhuman priority journal protozoon trypanosoma cruzi Animal Cattle Chromatography, Affinity Chromatography, DEAE-Cellulose Chromatography, Gel Cyclic AMP Immunoblotting Myocardium Phosphates Phosphorylation Protein Binding Protein Kinases Support, Non-U.S. Gov't Trypanosoma cruzi A cyclic AMP-dependent protein kinase activity from epimastigote forms of Trypanosoma cruzi was characterized. Cytosolic extracts were chromatographed on DEAE-cellulose columns, giving two peaks of kinase activity, which were eluted at 0.15 M- and 0.32 M-NaCl respectively. The second activity peak was stimulated by nanomolar concentrations of cyclic AMP. In addition, a cyclic AMP-binding protein co-eluted with the second kinase activity peak. Cyclic AMP-dependent protein kinase activity was further purified by gel filtration, affinity chromatography on histone-agarose and cyclic AMP-agarose, as well as by chromatography on CM-Sephadex. The enzyme ('holoenzyme') could be partially dissociated into two different components: 'catalytic' and 'regulatory'. The 'regulatory' component had specific binding for cyclic AMP, and it inhibited phosphotransferase activity of the homologous 'catalytic component' or of the 'catalytic subunit' from bovine heart. Cyclic AMP reversed these inhibitions. A 'holoenzyme preparation' was phosphorylated in the absence of exogenous phosphate acceptor and analyzed by polyacrylamide- gel electrophoresis. A 56 kDa band was phosphorylated. The same preparation was analysed by Western blotting, by using polyclonal antibodies to the regulatory subunits of protein kinases type I or II. Both antibodies reacted with the 56 kDa band. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_02646021_v255_n1_p319_Ulloa
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic antibody
cyclic amp
protein kinase
immunoblotting
nonhuman
priority journal
protozoon
trypanosoma cruzi
Animal
Cattle
Chromatography, Affinity
Chromatography, DEAE-Cellulose
Chromatography, Gel
Cyclic AMP
Immunoblotting
Myocardium
Phosphates
Phosphorylation
Protein Binding
Protein Kinases
Support, Non-U.S. Gov't
Trypanosoma cruzi
spellingShingle antibody
cyclic amp
protein kinase
immunoblotting
nonhuman
priority journal
protozoon
trypanosoma cruzi
Animal
Cattle
Chromatography, Affinity
Chromatography, DEAE-Cellulose
Chromatography, Gel
Cyclic AMP
Immunoblotting
Myocardium
Phosphates
Phosphorylation
Protein Binding
Protein Kinases
Support, Non-U.S. Gov't
Trypanosoma cruzi
Ulloa, R.M.
Mesri, E.
Esteva, M.
Torres, H.N.
Tellez-Inon, M.T.
Cyclic AMP-dependent protein kinase activity in Trypanosoma cruzi
topic_facet antibody
cyclic amp
protein kinase
immunoblotting
nonhuman
priority journal
protozoon
trypanosoma cruzi
Animal
Cattle
Chromatography, Affinity
Chromatography, DEAE-Cellulose
Chromatography, Gel
Cyclic AMP
Immunoblotting
Myocardium
Phosphates
Phosphorylation
Protein Binding
Protein Kinases
Support, Non-U.S. Gov't
Trypanosoma cruzi
description A cyclic AMP-dependent protein kinase activity from epimastigote forms of Trypanosoma cruzi was characterized. Cytosolic extracts were chromatographed on DEAE-cellulose columns, giving two peaks of kinase activity, which were eluted at 0.15 M- and 0.32 M-NaCl respectively. The second activity peak was stimulated by nanomolar concentrations of cyclic AMP. In addition, a cyclic AMP-binding protein co-eluted with the second kinase activity peak. Cyclic AMP-dependent protein kinase activity was further purified by gel filtration, affinity chromatography on histone-agarose and cyclic AMP-agarose, as well as by chromatography on CM-Sephadex. The enzyme ('holoenzyme') could be partially dissociated into two different components: 'catalytic' and 'regulatory'. The 'regulatory' component had specific binding for cyclic AMP, and it inhibited phosphotransferase activity of the homologous 'catalytic component' or of the 'catalytic subunit' from bovine heart. Cyclic AMP reversed these inhibitions. A 'holoenzyme preparation' was phosphorylated in the absence of exogenous phosphate acceptor and analyzed by polyacrylamide- gel electrophoresis. A 56 kDa band was phosphorylated. The same preparation was analysed by Western blotting, by using polyclonal antibodies to the regulatory subunits of protein kinases type I or II. Both antibodies reacted with the 56 kDa band.
format JOUR
author Ulloa, R.M.
Mesri, E.
Esteva, M.
Torres, H.N.
Tellez-Inon, M.T.
author_facet Ulloa, R.M.
Mesri, E.
Esteva, M.
Torres, H.N.
Tellez-Inon, M.T.
author_sort Ulloa, R.M.
title Cyclic AMP-dependent protein kinase activity in Trypanosoma cruzi
title_short Cyclic AMP-dependent protein kinase activity in Trypanosoma cruzi
title_full Cyclic AMP-dependent protein kinase activity in Trypanosoma cruzi
title_fullStr Cyclic AMP-dependent protein kinase activity in Trypanosoma cruzi
title_full_unstemmed Cyclic AMP-dependent protein kinase activity in Trypanosoma cruzi
title_sort cyclic amp-dependent protein kinase activity in trypanosoma cruzi
url http://hdl.handle.net/20.500.12110/paper_02646021_v255_n1_p319_Ulloa
work_keys_str_mv AT ulloarm cyclicampdependentproteinkinaseactivityintrypanosomacruzi
AT mesrie cyclicampdependentproteinkinaseactivityintrypanosomacruzi
AT estevam cyclicampdependentproteinkinaseactivityintrypanosomacruzi
AT torreshn cyclicampdependentproteinkinaseactivityintrypanosomacruzi
AT tellezinonmt cyclicampdependentproteinkinaseactivityintrypanosomacruzi
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