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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Detalles Bibliográficos
Publicado: 1988
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:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02646021_v255_n1_p319_Ulloa
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 paper:paper_02646021_v255_n1_p319_Ulloa
record_format dspace
spelling paper:paper_02646021_v255_n1_p319_Ulloa2025-07-30T18:01:23Z Cyclic AMP-dependent protein kinase activity in Trypanosoma cruzi 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. 1988 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02646021_v255_n1_p319_Ulloa 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
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.
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
publishDate 1988
url https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02646021_v255_n1_p319_Ulloa
http://hdl.handle.net/20.500.12110/paper_02646021_v255_n1_p319_Ulloa
_version_ 1840325121681653760

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