Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi

A Trypanosoma cruzi gene, PKB, coding for a putative protein kinase was cloned and sequenced. Analysis of the sequence showed that the encoded protein (called PKB) corresponds to a relatively novel subgroup of Ser/Thr protein kinases denominated protein kinases B (PKB), related to A and C protein ki...

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Autores principales:	Pascuccelli, V., Labriola, C., Téllez-Iñón, M.T., Parodi, A.J.
Formato:	JOUR
Materias:	Akt protein kinase Protein kinase B RAK protein kinase Trypanosoma cruzi protein kinase amino acid sequence animal experiment article controlled study epimastigote gene expression nonhuman nucleotide sequence polyacrylamide gel electrophoresis priority journal protein analysis signal transduction trypanosoma cruzi trypomastigote western blotting Amino Acid Sequence Animals Blotting, Western Cloning, Molecular DNA, Protozoan Escherichia coli Hydrogen-Ion Concentration Molecular Sequence Data Phosphorylation Polymerase Chain Reaction Protein-Serine-Threonine Kinases Proto-Oncogene Proteins Proto-Oncogene Proteins c-akt Recombinant Proteins Sequence Analysis, DNA Temperature Mammalia Protozoa Trypanosoma Trypanosomatidae unidentified retrovirus
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_01666851_v102_n1_p21_Pascuccelli
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	todo:paper_01666851_v102_n1_p21_Pascuccelli2023-10-03T15:04:04Z Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi Pascuccelli, V. Labriola, C. Téllez-Iñón, M.T. Parodi, A.J. Akt protein kinase Protein kinase B RAK protein kinase Trypanosoma cruzi protein kinase amino acid sequence animal experiment article controlled study epimastigote gene expression nonhuman nucleotide sequence polyacrylamide gel electrophoresis priority journal protein analysis signal transduction trypanosoma cruzi trypomastigote western blotting Amino Acid Sequence Animals Blotting, Western Cloning, Molecular DNA, Protozoan Escherichia coli Hydrogen-Ion Concentration Molecular Sequence Data Phosphorylation Polymerase Chain Reaction Protein-Serine-Threonine Kinases Proto-Oncogene Proteins Proto-Oncogene Proteins c-akt Recombinant Proteins Sequence Analysis, DNA Temperature Trypanosoma cruzi Escherichia coli Mammalia Protozoa Trypanosoma Trypanosoma cruzi Trypanosomatidae unidentified retrovirus A Trypanosoma cruzi gene, PKB, coding for a putative protein kinase was cloned and sequenced. Analysis of the sequence showed that the encoded protein (called PKB) corresponds to a relatively novel subgroup of Ser/Thr protein kinases denominated protein kinases B (PKB), related to A and C protein kinases (RAC), or protein kinases of the transforming retrovirus AKT8 (Akt) in which the catalytic domains show similarity to corresponding domains of protein kinases A and protein kinases C. Unlike mammalian enzymes belonging to the same subgroup, PKB did not have a pleckstrin (PH)-homologous domain. PKB was expressed in Escherichia coli and the recombinant protein was found to be a Thr-specific protein kinase that required Mn2+ for activity and used ATP as phosphate donor (K(m)=1.8 μM). Classical protein kinase A and protein kinase C modulators and inhibitors were found to have only marginal or no effect on PKB activity. Antisera raised against the recombinant protein recognized PKB in Western blotting analysis of cell extracts as a membrane bound protein. Evidence was obtained suggesting the presence of a Cys-linked acyl anchor. Northern and Western blotting analysis showed that PKB was constitutively expressed in the lag, exponential and stationary phases of T. cruzi epimastigote growth, as well as in the amastigote and metacyclic trypomastigote stages of differentiation. This is the first description of the existence of a protein kinase B in trypanosomatid protozoa. Copyright (C) 1999 Elsevier Science B.V. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_01666851_v102_n1_p21_Pascuccelli
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Akt protein kinase Protein kinase B RAK protein kinase Trypanosoma cruzi protein kinase amino acid sequence animal experiment article controlled study epimastigote gene expression nonhuman nucleotide sequence polyacrylamide gel electrophoresis priority journal protein analysis signal transduction trypanosoma cruzi trypomastigote western blotting Amino Acid Sequence Animals Blotting, Western Cloning, Molecular DNA, Protozoan Escherichia coli Hydrogen-Ion Concentration Molecular Sequence Data Phosphorylation Polymerase Chain Reaction Protein-Serine-Threonine Kinases Proto-Oncogene Proteins Proto-Oncogene Proteins c-akt Recombinant Proteins Sequence Analysis, DNA Temperature Trypanosoma cruzi Escherichia coli Mammalia Protozoa Trypanosoma Trypanosoma cruzi Trypanosomatidae unidentified retrovirus
spellingShingle	Akt protein kinase Protein kinase B RAK protein kinase Trypanosoma cruzi protein kinase amino acid sequence animal experiment article controlled study epimastigote gene expression nonhuman nucleotide sequence polyacrylamide gel electrophoresis priority journal protein analysis signal transduction trypanosoma cruzi trypomastigote western blotting Amino Acid Sequence Animals Blotting, Western Cloning, Molecular DNA, Protozoan Escherichia coli Hydrogen-Ion Concentration Molecular Sequence Data Phosphorylation Polymerase Chain Reaction Protein-Serine-Threonine Kinases Proto-Oncogene Proteins Proto-Oncogene Proteins c-akt Recombinant Proteins Sequence Analysis, DNA Temperature Trypanosoma cruzi Escherichia coli Mammalia Protozoa Trypanosoma Trypanosoma cruzi Trypanosomatidae unidentified retrovirus Pascuccelli, V. Labriola, C. Téllez-Iñón, M.T. Parodi, A.J. Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi
topic_facet	Akt protein kinase Protein kinase B RAK protein kinase Trypanosoma cruzi protein kinase amino acid sequence animal experiment article controlled study epimastigote gene expression nonhuman nucleotide sequence polyacrylamide gel electrophoresis priority journal protein analysis signal transduction trypanosoma cruzi trypomastigote western blotting Amino Acid Sequence Animals Blotting, Western Cloning, Molecular DNA, Protozoan Escherichia coli Hydrogen-Ion Concentration Molecular Sequence Data Phosphorylation Polymerase Chain Reaction Protein-Serine-Threonine Kinases Proto-Oncogene Proteins Proto-Oncogene Proteins c-akt Recombinant Proteins Sequence Analysis, DNA Temperature Trypanosoma cruzi Escherichia coli Mammalia Protozoa Trypanosoma Trypanosoma cruzi Trypanosomatidae unidentified retrovirus
description	A Trypanosoma cruzi gene, PKB, coding for a putative protein kinase was cloned and sequenced. Analysis of the sequence showed that the encoded protein (called PKB) corresponds to a relatively novel subgroup of Ser/Thr protein kinases denominated protein kinases B (PKB), related to A and C protein kinases (RAC), or protein kinases of the transforming retrovirus AKT8 (Akt) in which the catalytic domains show similarity to corresponding domains of protein kinases A and protein kinases C. Unlike mammalian enzymes belonging to the same subgroup, PKB did not have a pleckstrin (PH)-homologous domain. PKB was expressed in Escherichia coli and the recombinant protein was found to be a Thr-specific protein kinase that required Mn2+ for activity and used ATP as phosphate donor (K(m)=1.8 μM). Classical protein kinase A and protein kinase C modulators and inhibitors were found to have only marginal or no effect on PKB activity. Antisera raised against the recombinant protein recognized PKB in Western blotting analysis of cell extracts as a membrane bound protein. Evidence was obtained suggesting the presence of a Cys-linked acyl anchor. Northern and Western blotting analysis showed that PKB was constitutively expressed in the lag, exponential and stationary phases of T. cruzi epimastigote growth, as well as in the amastigote and metacyclic trypomastigote stages of differentiation. This is the first description of the existence of a protein kinase B in trypanosomatid protozoa. Copyright (C) 1999 Elsevier Science B.V.
format	JOUR
author	Pascuccelli, V. Labriola, C. Téllez-Iñón, M.T. Parodi, A.J.
author_facet	Pascuccelli, V. Labriola, C. Téllez-Iñón, M.T. Parodi, A.J.
author_sort	Pascuccelli, V.
title	Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi
title_short	Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi
title_full	Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi
title_fullStr	Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi
title_full_unstemmed	Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi
title_sort	molecular and biochemical characterization of a protein kinase b from trypanosoma cruzi
url	http://hdl.handle.net/20.500.12110/paper_01666851_v102_n1_p21_Pascuccelli
work_keys_str_mv	AT pascuccelliv molecularandbiochemicalcharacterizationofaproteinkinasebfromtrypanosomacruzi AT labriolac molecularandbiochemicalcharacterizationofaproteinkinasebfromtrypanosomacruzi AT tellezinonmt molecularandbiochemicalcharacterizationofaproteinkinasebfromtrypanosomacruzi AT parodiaj molecularandbiochemicalcharacterizationofaproteinkinasebfromtrypanosomacruzi
_version_	1807322047078989824

Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi

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