title: Immunological identification of a cAMP-dependent protein kinase regulatory subunit-like protein from the Trypanosoma equiperdum TeAp-N/D1 isolate.
abstract: Polyclonal immunoglobulin Y (IgY) antibodies were produced in chicken eggs against the purified R(II)-subunit of the cAMP-dependent protein kinase (PKA) from pig heart, which corresponds to the Sus scrofa R(II)α isoform. In order to evaluate whether Trypanosoma equiperdum possessed PKA R-like proteins, parasites from the Venezuelan TeAp-N/D1 strain were examined using the generated anti-R(II) IgY antibodies. Western blot experiments revealed a 57-kDa polypeptide band that was distinctively recognized by these antibodies. Likewise, polyclonal antibodies raised in mice ascites against the recombinant T. equiperdum PKA R-like protein recognized the PKA R(II)-subunit purified from porcine heart and the recombinant human PKA R(I)β-subunit by immunoblotting. However, a partially purified fraction of the parasite PKA R-like protein was not capable of binding cAMP, implying that this protein is not a direct downstream cAMP effector in T. equiperdum. Although the function of the S. scrofa PKA R(II)α and the T. equiperdum PKA R-like protein appear to be different, their cross-reactivity together with results obtained by bioinformatics techniques corroborated the high level of homology exhibited by both proteins. Moreover, its presence in other trypanosomatids suggests an important cellular role of PKA R-like proteins in parasite physiology.
authors.affiliation: a Departamento de Biología Celular , Universidad Simón Bolívar , Caracas , Venezuela.
authors.forename: Maritza
authors.initials: M
authors.lastname: Calabokis
authors.affiliation: a Departamento de Biología Celular , Universidad Simón Bolívar , Caracas , Venezuela.|b Licenciatura en Química , Universidad Simón Bolívar , Caracas , Venezuela.
authors.forename: Yelvis
authors.initials: Y
authors.lastname: González
authors.affiliation: a Departamento de Biología Celular , Universidad Simón Bolívar , Caracas , Venezuela.|b Licenciatura en Química , Universidad Simón Bolívar , Caracas , Venezuela.
authors.forename: Adriana
authors.initials: A
authors.lastname: Merchán
authors.affiliation: a Departamento de Biología Celular , Universidad Simón Bolívar , Caracas , Venezuela.|c Postgrado en Química , Universidad Simón Bolívar , Caracas , Venezuela.
authors.forename: José L
authors.initials: JL
authors.lastname: Escalona
authors.affiliation: a Departamento de Biología Celular , Universidad Simón Bolívar , Caracas , Venezuela.|c Postgrado en Química , Universidad Simón Bolívar , Caracas , Venezuela.
authors.forename: Nelson A
authors.initials: NA
authors.lastname: Araujo
authors.affiliation: a Departamento de Biología Celular , Universidad Simón Bolívar , Caracas , Venezuela.|d Postgrado en Ciencias Biológicas , Universidad Simón Bolívar , Caracas , Venezuela.|e Laboratorio de Dinámica Estocástica, Centro de Física , Instituto Venezolano de Investigaciones Científicas , Caracas , Venezuela.
authors.forename: Carlos E
authors.initials: CE
authors.lastname: Sanz-Rodríguez
authors.affiliation: a Departamento de Biología Celular , Universidad Simón Bolívar , Caracas , Venezuela.|d Postgrado en Ciencias Biológicas , Universidad Simón Bolívar , Caracas , Venezuela.
authors.forename: Carolina
authors.initials: C
authors.lastname: Cywiak
authors.affiliation: a Departamento de Biología Celular , Universidad Simón Bolívar , Caracas , Venezuela.|f Departamento de Biología , Universidad de Yachay Tech , Hacienda San José , Urcuquí , Ecuador.
authors.forename: Lilian M
authors.initials: LM
authors.lastname: Spencer
authors.affiliation: g Dirección de Salud , Fundación Instituto de Estudios Avanzados IDEA , Caracas , Venezuela.
authors.forename: Juan C
authors.initials: JC
authors.lastname: Martínez
authors.affiliation: a Departamento de Biología Celular , Universidad Simón Bolívar , Caracas , Venezuela.
authors.forename: José
authors.initials: J
authors.lastname: Bubis
mesh_terms: D000818:Animals; D002645:Chickens; D017868:Cyclic AMP-Dependent Protein Kinases / Q000032:analysis* / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007136:Immunoglobulins / Q000276:immunology; D051379:Mice; D021122:Protein Subunits / Q000032:analysis* / Q000276:immunology* / Q000378:metabolism; D014345:Trypanosoma / Q000201:enzymology* / Q000276:immunology / Q000302:isolation & purification
chemical_list: C016512:IgY; D007136:Immunoglobulins; D021122:Protein Subunits; D017868:Cyclic AMP-Dependent Protein Kinases
keywords: Trypanosoma equiperdum; cAMP; immunoglobulin Y antibodies; immunological cross-reactivity; regulatory subunits of the cAMP-dependent protein kinase