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Abstract:

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.

Registro:

Documento: Artículo
Título:Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi
Autor:Pascuccelli, V.; Labriola, C.; Téllez-Iñón, M.T.; Parodi, A.J.
Filiación:Inst. Invest. Bioquimicas Fund. C., Antonio Machado 151, 1405, Buenos Aires, Argentina
Inst. Ing. Genet. Y Biol. Molec. (I., Vuelta de Obligado 2490, 1428, Buenos Aires, Argentina
Palabras clave: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
Año:1999
Volumen:102
Número:1
Página de inicio:21
Página de fin:33
DOI: http://dx.doi.org/10.1016/S0166-6851(99)00076-6
Título revista:Molecular and Biochemical Parasitology
Título revista abreviado:Mol. Biochem. Parasitol.
ISSN:01666851
CODEN:MBIPD
CAS:DNA, Protozoan; Protein-Serine-Threonine Kinases, EC 2.7.1.37; Proto-Oncogene Proteins c-akt, EC 2.7.1.37; Proto-Oncogene Proteins; Recombinant Proteins
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v102_n1_p21_Pascuccelli

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Citas:

---------- APA ----------
Pascuccelli, V., Labriola, C., Téllez-Iñón, M.T. & Parodi, A.J. (1999) . Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi. Molecular and Biochemical Parasitology, 102(1), 21-33.
http://dx.doi.org/10.1016/S0166-6851(99)00076-6
---------- CHICAGO ----------
Pascuccelli, V., Labriola, C., Téllez-Iñón, M.T., Parodi, A.J. "Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi" . Molecular and Biochemical Parasitology 102, no. 1 (1999) : 21-33.
http://dx.doi.org/10.1016/S0166-6851(99)00076-6
---------- MLA ----------
Pascuccelli, V., Labriola, C., Téllez-Iñón, M.T., Parodi, A.J. "Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi" . Molecular and Biochemical Parasitology, vol. 102, no. 1, 1999, pp. 21-33.
http://dx.doi.org/10.1016/S0166-6851(99)00076-6
---------- VANCOUVER ----------
Pascuccelli, V., Labriola, C., Téllez-Iñón, M.T., Parodi, A.J. Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi. Mol. Biochem. Parasitol. 1999;102(1):21-33.
http://dx.doi.org/10.1016/S0166-6851(99)00076-6