title: Identification of Besnoitia besnoiti proteins that showed differences in abundance between tachyzoite and bradyzoite stages by difference gel electrophoresis.
abstract: Bovine besnoitiosis is a chronic and debilitating disease, caused by the apicomplexan parasite Besnoitia besnoiti. Infection of cattle by B. besnoiti is governed by the tachyzoite stage, which is related to acute infection, and the bradyzoite stage gathered into macroscopic cysts located in subcutaneous tissue in the skin, mucosal membranes and sclera conjunctiva and related to persistence and chronic infection. However, the entire life cycle of this parasite and the molecular mechanisms underlying tachyzoite-to-bradyzoite conversion remain unknown. In this context, a different antigenic pattern has been observed between tachyzoite and bradyzoite extracts. Thus, to identify stage-specific proteins, a difference gel electrophoresis (DIGE) approach was used on tachyzoite and bradyzoite extracts followed by mass spectrometry (MS) analysis. A total of 130 and 132 spots were differentially expressed in bradyzoites and tachyzoites, respectively (average ratio ± 1.5, P<0.05 in t-test). Furthermore, 25 differentially expressed spots were selected and analysed by MALDI-TOF/MS. As a result, 5 up-regulated bradyzoite proteins (GAPDH, ENO1, LDH, SOD and RNA polymerase) and 5 up-regulated tachyzoite proteins (ENO2; LDH; ATP synthase; HSP70 and PDI) were identified. The present results set the basis for the identification of new proteins as drug targets. Moreover, the role of these proteins in tachyzoite-to-bradyzoite conversion and the role of the host cell environment should be a subject of further research.
authors.affiliation: SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040-Madrid, Spain.
authors.forename: Aurora
authors.initials: A
authors.lastname: Fernández-García
authors.forename: Gema
authors.initials: G
authors.lastname: Alvarez-García
authors.forename: Virginia
authors.initials: V
authors.lastname: Marugán-Hernández
authors.forename: Paula
authors.initials: P
authors.lastname: García-Lunar
authors.forename: Adriana
authors.initials: A
authors.lastname: Aguado-Martínez
authors.forename: Verónica
authors.initials: V
authors.lastname: Risco-Castillo
authors.forename: Luis M
authors.initials: LM
authors.lastname: Ortega-Mora
mesh_terms: D000818:Animals; D002417:Cattle; D003048:Coccidiosis / Q000469:parasitology / Q000662:veterinary*; D005786:Gene Expression Regulation; D008018:Life Cycle Stages; D040901:Proteomics; D015800:Protozoan Proteins / Q000032:analysis / Q000378:metabolism*; D021745:Sarcocystidae / Q000737:chemistry / Q000254:growth & development* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000662:veterinary; D057825:Two-Dimensional Difference Gel Electrophoresis / Q000662:veterinary
chemical_list: D015800:Protozoan Proteins