Characterization of the catalytic subunit of Trypanosoma cruzi cyclic AMP-dependent protein kinase

The catalytic subunit of cyclic AMP-dependent protein kinase from Trypanosoma cruzi epimastigote forms was purified by ionic-exchange chromatography, affinity chromatography and sucrose gradient centrifugation. Upon polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate, the p...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Ochatt, Claudia Margarita, Torres, Héctor Norberto
Publicado: 1993
Materias:
Catalytic subunit
Cyclic AMP
Protein kinase A
Trypanosoma cruzi
cyclic amp
protein kinase
article
catalysis
enzyme activity
enzyme analysis
nonhuman
priority journal
trypanosoma cruzi
Animal
Binding Sites
Kinetics
Molecular Structure
Molecular Weight
Protein Conformation
Protein Kinases
Support, Non-U.S. Gov't
Acceso en línea:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v57_n1_p73_Ochatt
http://hdl.handle.net/20.500.12110/paper_01666851_v57_n1_p73_Ochatt
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
id paper:paper_01666851_v57_n1_p73_Ochatt
record_format dspace
spelling paper:paper_01666851_v57_n1_p73_Ochatt2023-06-08T15:16:06Z Characterization of the catalytic subunit of Trypanosoma cruzi cyclic AMP-dependent protein kinase Ochatt, Claudia Margarita Torres, Héctor Norberto Catalytic subunit Cyclic AMP Protein kinase A Trypanosoma cruzi cyclic amp protein kinase article catalysis enzyme activity enzyme analysis nonhuman priority journal trypanosoma cruzi Animal Binding Sites Cyclic AMP Kinetics Molecular Structure Molecular Weight Protein Conformation Protein Kinases Support, Non-U.S. Gov't Trypanosoma cruzi The catalytic subunit of cyclic AMP-dependent protein kinase from Trypanosoma cruzi epimastigote forms was purified by ionic-exchange chromatography, affinity chromatography and sucrose gradient centrifugation. Upon polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate, the purified preparations showed a main polypeptide band with a mobility of about 40 kDa. In Western blots this band immunoreacted with a polyclonal antibody specific for the catalytic subunit of bovine heart protein kinase A. Hydrodynamic and molecular parameters of this subunit are as follows: molecular weight, 40 000 ± 3000; sedimentation constant, 2.8 ± 0.3 S; Stokes' radius, 2.8 ± 0.2 nm; frictional ratio, 1.28 ± 0.05. Purified preparations of T. cruzi catalytic and regulatory subunits reconstitute a holoenzyme with a sedimentation constant, 8.6 ± 1.17 S. This data together with those previously reported by Ulloa et al. [8] indicate that the T. cruzi cyclic AMP-dependent protein kinase holoenzyme is a tetramer with the structure R2C2 of about 200 kDa. The apparent Km of the catalytic subunit for ATP and histone IIA or kemptide as phosphate donor and acceptor, respectively, were 40 μM, 48.6 μM and 26 μM, respectively. © 1993. Fil:Ochatt, C.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Torres, H.N. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1993 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v57_n1_p73_Ochatt http://hdl.handle.net/20.500.12110/paper_01666851_v57_n1_p73_Ochatt
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic Catalytic subunit
Cyclic AMP
Protein kinase A
Trypanosoma cruzi
cyclic amp
protein kinase
article
catalysis
enzyme activity
enzyme analysis
nonhuman
priority journal
trypanosoma cruzi
Animal
Binding Sites
Cyclic AMP
Kinetics
Molecular Structure
Molecular Weight
Protein Conformation
Protein Kinases
Support, Non-U.S. Gov't
Trypanosoma cruzi
spellingShingle Catalytic subunit
Cyclic AMP
Protein kinase A
Trypanosoma cruzi
cyclic amp
protein kinase
article
catalysis
enzyme activity
enzyme analysis
nonhuman
priority journal
trypanosoma cruzi
Animal
Binding Sites
Cyclic AMP
Kinetics
Molecular Structure
Molecular Weight
Protein Conformation
Protein Kinases
Support, Non-U.S. Gov't
Trypanosoma cruzi
Ochatt, Claudia Margarita
Torres, Héctor Norberto
Characterization of the catalytic subunit of Trypanosoma cruzi cyclic AMP-dependent protein kinase
topic_facet Catalytic subunit
Cyclic AMP
Protein kinase A
Trypanosoma cruzi
cyclic amp
protein kinase
article
catalysis
enzyme activity
enzyme analysis
nonhuman
priority journal
trypanosoma cruzi
Animal
Binding Sites
Cyclic AMP
Kinetics
Molecular Structure
Molecular Weight
Protein Conformation
Protein Kinases
Support, Non-U.S. Gov't
Trypanosoma cruzi
description The catalytic subunit of cyclic AMP-dependent protein kinase from Trypanosoma cruzi epimastigote forms was purified by ionic-exchange chromatography, affinity chromatography and sucrose gradient centrifugation. Upon polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate, the purified preparations showed a main polypeptide band with a mobility of about 40 kDa. In Western blots this band immunoreacted with a polyclonal antibody specific for the catalytic subunit of bovine heart protein kinase A. Hydrodynamic and molecular parameters of this subunit are as follows: molecular weight, 40 000 ± 3000; sedimentation constant, 2.8 ± 0.3 S; Stokes' radius, 2.8 ± 0.2 nm; frictional ratio, 1.28 ± 0.05. Purified preparations of T. cruzi catalytic and regulatory subunits reconstitute a holoenzyme with a sedimentation constant, 8.6 ± 1.17 S. This data together with those previously reported by Ulloa et al. [8] indicate that the T. cruzi cyclic AMP-dependent protein kinase holoenzyme is a tetramer with the structure R2C2 of about 200 kDa. The apparent Km of the catalytic subunit for ATP and histone IIA or kemptide as phosphate donor and acceptor, respectively, were 40 μM, 48.6 μM and 26 μM, respectively. © 1993.
author Ochatt, Claudia Margarita
Torres, Héctor Norberto
author_facet Ochatt, Claudia Margarita
Torres, Héctor Norberto
author_sort Ochatt, Claudia Margarita
title Characterization of the catalytic subunit of Trypanosoma cruzi cyclic AMP-dependent protein kinase
title_short Characterization of the catalytic subunit of Trypanosoma cruzi cyclic AMP-dependent protein kinase
title_full Characterization of the catalytic subunit of Trypanosoma cruzi cyclic AMP-dependent protein kinase
title_fullStr Characterization of the catalytic subunit of Trypanosoma cruzi cyclic AMP-dependent protein kinase
title_full_unstemmed Characterization of the catalytic subunit of Trypanosoma cruzi cyclic AMP-dependent protein kinase
title_sort characterization of the catalytic subunit of trypanosoma cruzi cyclic amp-dependent protein kinase
publishDate 1993
url https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v57_n1_p73_Ochatt
http://hdl.handle.net/20.500.12110/paper_01666851_v57_n1_p73_Ochatt
work_keys_str_mv AT ochattclaudiamargarita characterizationofthecatalyticsubunitoftrypanosomacruzicyclicampdependentproteinkinase
AT torreshectornorberto characterizationofthecatalyticsubunitoftrypanosomacruzicyclicampdependentproteinkinase
_version_ 1768542641383800832

Ejemplares similares