Archives of Microbiology

, Volume 187, Issue 1, pp 55–66 | Cite as

Protein kinase C isoforms from Giardia duodenalis: identification and functional characterization of a β-like molecule during encystment

  • María Luisa Bazán-Tejeda
  • Raúl Argüello-García
  • Rosa María Bermúdez-Cruz
  • Martha Robles-Flores
  • Guadalupe Ortega-Pierres
Original Paper

Abstract

Protein kinase C (PKC) is a family of serine/threonine kinases that regulate many different cellular processes such as cell growth and differentiation in eukaryotic cells. Using specific polyclonal antibodies raised against mammalian PKC isoforms, it was demonstrated here for the first time that Giardia duodenalis expresses several PKC isoforms (beta, delta, epsilon, theta and zeta). All PKC isoforms detected showed changes in their expression pattern during encystment induction. In addition, selective PKC inhibitors blocked the encystment in a dose-dependent manner, suggesting that PKC isozymes may play important roles during this differentiation process. We have characterized here the only conventional-type PKC member found so far in Giardia, which showed an increased expression and changes in its intracellular localization pattern during cyst formation. The purified protein obtained by chromatography on DEAE-cellulose followed by size-exclusion chromatography, displayed in vitro kinase activity using histone HI-IIIS as substrate, which was dependent on cofactors required by conventional PKCs, i.e., phospholipids and calcium. An open reading frame in the Giardia Genome Database that encodes a homolog of PKCβ catalytic domain was identified and cloned. The expressed recombinant protein was also recognized by a mammalian anti-PKCβ antibody and was referred as giardial PKCβ on the basis of all these experimental evidence.

Keywords

Giardia Protein kinase C Encystment Phosphorylation Translocation Lower eukaryotes Protozoan differentiation 

Abbreviations

CWP

Cell wall protein

PKA

Protein kinase A

DAG

Diacylglycerol

IPTG

Isopropyl-β-d-thiogalactopyranoside

BIM

Bisindolylmaleimide

ORF

Open reading frame

PS

Phosphatidylserine

cPKC

Conventional protein kinase C

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • María Luisa Bazán-Tejeda
    • 1
  • Raúl Argüello-García
    • 1
  • Rosa María Bermúdez-Cruz
    • 1
  • Martha Robles-Flores
    • 2
  • Guadalupe Ortega-Pierres
    • 1
  1. 1.Departamento de Genética y Biología MolecularCentro de Investigación y de Estudios Avanzados-IPN (CINVESTAV)Mexico D.F.Mexico
  2. 2.Departamento de Bioquímica, Facultad de MedicinaUniversidad Nacional Autónoma de MéxicoMexico D.F.Mexico

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