Journal of Proteomics & Bioinformatics
Open Access
ISSN: 0974-276X
ISSN: 0974-276X
Pedro J. Alcolea, Ana Alonso and Vicente Larraga
Posters: J Proteomics Bioinform
The genus Crithidia comprises monoxenic parasite species that infect diverse genera of hexapods such Diptera and Hymenoptera. Their digenetic counterparts Trypanosoma spp. and Leishmania spp. are respectively responsible for sleeping sickness, Chagas? disease and leishmaniasis. The genome of Crithidia has not been still sequenced unlike those of T. cruzi, T. brucei and three species of the genus Leishmania. In addition, little is known about differential gene expression of Crithidia spp. The aim of this research is the detection of differentially regulated proteins in choanomastigotes of C. fasciculata. We performed triplicate cultures monitoring the growth curves reaching the stationary phase in four days. Total protein extracts were prepared daily, precipitated with trichloroacetic acid and quantified by the bicinchoninic acid method. 50μg of each extract were 2D electrophoresed (isoelectrofocusing in pH 3 to 10 non-linear gradient), stained with SYPRO Ruby and analyzed with the PD-QUEST software. The normalization algorithm was total quantity in valid spots and significant differences inferred by the paired t-test. Protein identification was carried out by MALDI-TOF/TOF mass spectrometry. Regarding the results, glycolysis is activated in logarithmic phase, as the aldolase, the phosphoglycerate mutase, the enolase and the glycosomal malate dehydrogenase are over-expressed. We reported the same in Leishmania infantum, the etiological agent of visceral leishmaniasis in the Mediterranean basin. Finally, the tryparedoxin peroxidase and the trypanothione reductase are up-regulated in stationary phase C. fasciculata choanomastigotes, which suggests a progressive increase of oxidative stress and is indicative of changes in redox homeostasis.
Pedro J. Alcolea is PhD in Biology by the Complutense University of Madrid (Spain) and staff member of the CIB-CSIC Molecular Parasitology group. Ana Alonso is PhD in Veterinary Science by the Complutense University of Madrid and staff member of the CIB-CSIC Molecular Parasitology group. Vicente Larraga is MD and PhD in Biology by the Complutense University of Madrid and staff Research Professor of the CIB-CSIC. He is the Scientific Responsible of the CIB Molecular Parasitology group.