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Tight binding between a pool of the heterodimeric α/β tubulin and a protein kinase CK2 in Trypanosoma cruzi epimastigotes

Published online by Cambridge University Press:  07 December 2005

A. R. DE LIMA ,
R. MEDINA ,
G. L. UZCANGA ,
K. NORIS SUÁREZ ,
V. T. CONTRERAS ,
M. C. NAVARRO ,
R. ARTEAGA  and
J. BUBIS
A. R. DE LIMA
Affiliation:
Departamento de Biología Celular, Universidad Simón Bolívar, Apartado 89.000, Valle de Sartenejas, Caracas 1081-A, Venezuela Laboratorio de Protozoología, Centro BioMolP, Facultad de Ciencias de la Salud, Universidad de Carabobo, Valencia, Venezuela
R. MEDINA
Affiliation:
Departamento de Biología Celular, Universidad Simón Bolívar, Apartado 89.000, Valle de Sartenejas, Caracas 1081-A, Venezuela
G. L. UZCANGA
Affiliation:
Departamento de Biología Celular, Universidad Simón Bolívar, Apartado 89.000, Valle de Sartenejas, Caracas 1081-A, Venezuela
K. NORIS SUÁREZ
Affiliation:
Departamento de Biología Celular, Universidad Simón Bolívar, Apartado 89.000, Valle de Sartenejas, Caracas 1081-A, Venezuela
V. T. CONTRERAS
Affiliation:
Laboratorio de Protozoología, Centro BioMolP, Facultad de Ciencias de la Salud, Universidad de Carabobo, Valencia, Venezuela
M. C. NAVARRO
Affiliation:
Laboratorio de Protozoología, Centro BioMolP, Facultad de Ciencias de la Salud, Universidad de Carabobo, Valencia, Venezuela
R. ARTEAGA
Affiliation:
Laboratorio de Protozoología, Centro BioMolP, Facultad de Ciencias de la Salud, Universidad de Carabobo, Valencia, Venezuela
J. BUBIS
Affiliation:
Departamento de Biología Celular, Universidad Simón Bolívar, Apartado 89.000, Valle de Sartenejas, Caracas 1081-A, Venezuela
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Abstract

Tubulin is the predominant phosphoprotein in Trypanosoma cruzi epimastigotes and is phosphorylated by a protein kinase CK2. Interestingly, the presence or absence of divalent cations affected the solubilization of a pool of the parasite tubulin and the CK2 responsible for its phosphorylation. This fraction of tubulin and its kinase co-eluted using phosphocellulose, DEAE-Sepharose and Sephacryl S-300 chromatographies. Anti-α tubulin antibodies co-immunoprecipitated both tubulin and the CK2 responsible for its phosphorylation, and anti-CK2 α-subunit antibodies immunoprecipitated radioactively labelled α and β tubulin from phosphorylated epimastigote homogenates. Additionally, native polyacrylamide gel electrophoresis of the purified and radioactively labelled fraction containing tubulin and its kinase demonstrated the phosphorylation of a unique band that reacted with both anti-CK2 α-subunit and anti-tubulin antibodies. Together, these results establish a strong interaction between a pool of the heterodimeric α/β tubulin and a CK2 in this parasite. Hydrodynamic measurements indicated that the T. cruzi tubulin-CK2 complex is globular with an estimated size of 145·4–147·5 kDa.

Keywords

cytoskeletonmicrotubulesprotein kinase CK2signal transductionTrypanosoma cruzi epimastigotestubulin
Type
Research Article
Information
Parasitology , Volume 132 , Issue 4 , April 2006 , pp. 511 - 523
Copyright
2005 Cambridge University Press

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