Bernal, Dolores

Esteban, José Guillermo

Marcilla, Antonio

Pérez-García, Ana

Sotillo, Javier

Sánchez del Pino, Manuel M.

Toledo, Rafael

Valero, L.

Repositorio Institucional de la Universidad de Córdoba

S6. Animal, Plant and Microbial Proteomics Posters164P. 92ENOLASE IN THE INTESTINAL HELMINTH INFECTIONS: THE CASE OF THE ECHiNOSTOMA CAPRONi-MICE MODELSotillo J.1, Pérez-García A.1, Bernal D.2, Sanchez del Pino M.M.3, Valero L.3, Esteban J.G.1, Marcilla A.1, Toledo R.11Dep. Biología Celular y Parasitología, Facultat de Farmàcia, Universitat de Valencia, Valencia, Spain; 2Laboratorio de Proteómica, Centro de Investigación Principe Felipe, Valencia, Spain; 3Dep. Bioquímica y Biología Molecular, Facultat de Ciències Biològiques, Universitat de València, Valencia, Spain.Echinostomes are intestinal parasitic trematodes which do not migrate within the tissue of their definitive hosts. These parasites have been extensively used as experimental models for the study of the host-parasite relationships. Our group has employed the model Echinostoma caproni (Trematoda: Echinostomatidae)-rodent to gain further insight in the molecules involved in the host-parasite interface that may be of importance to determine the course of the infection. This is based on the observation that E. caproni develops chronic or acute infections depending on the rodent host used (mice or rat, respectively). In the present study, we identify the enolase as the most immunogenic molecule of the excretory-secretory products (ESP) of E. caproni infected mice and we also report on the molecular cloning and characterization of this protein.Antigenic proteins of E. caproni ESP were investigated by immunoproteomics. ESP of E. caproni separated by 2-D gel electrophoresis were transferred to nitrocellulose membranes and probed with different mice immunoglobulin classes. Enolase was recognized in 8 different spots of which 7 of them were detected in the expected molecular weight and were recognized by IgA, IgG or IgG and IgG1. Another spot identified as enolase a 72 kDa was recognized by IgM. Digestion with n-glycosidase F rendered a polypeptide with a molecular weight similar to that expected for enolase. Molecular cloning and in vitro expression in Escherichia coli of E. caproni enolase allowed us to determine that the protein contains 431 aminoacids and a theoretical MW of 46 kDa. The recombinant protein binds specifically to human plasminogen in vitro, confirming its properties as a host-interacting protein.Acknowledgements: This study was supported by the project CGL2005-02321/BOS from the Ministerio de Educación y Ciencia (Spain) and FEDER (EU), the projects GV07/006, GVPRE/2008/049 and complementary grant ACOMP07/071 from the Conselleria d’Empresa, Universitat I Ciència de la Generalitat Valenciana (Spain). This work has been carried out while the first author (J.S.) was recipient of a predoctoral fellowship from the Ministerio de Educación y Ciencia, Madrid (Spain) and the second author (A.P.) was recipient of a fellowship (BC06-284) from the Universitat de València, funded by DGSP (185/2006).

Enolase in the intestinal helminth infections: the case of the echinostoma caproni-mice model

Enolase in the intestinal helminth infections: the case of the echinostoma caproni-mice model

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Repositorio Institucional de la Universidad de Córdoba