A new real-time RT-PCR method allowing reliable absolute quantification of human cytokine mRNA in paediatric malaria clinical samples |
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| Authors: | P BOEUF I VIGAN D JUBLOT J-C BARALE B GOKA J KURTZHALS B D AKANMORI L HVIID O PUIJALON C BEHR |
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| Institution: | Laboratoire de Biologie Fonctionnelle des Protozoaires, USM504, Departement Regulations, Developpement, Diversite Moleculaire, Museum National d'Histoire Naturelle, 61 rue Buffon, 75231 Paris Cedex 05, France,;Laboratorio Multidisciplinar de Pesquisa em Doença de Chagas, CP04536, Universidade de Brasilia, Brasilia, Brazil |
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| Abstract: | Proteases play important roles in several processes of the biology of parasites including interactions with their hosts. We have reported that prolyl oligopeptidase from Trypanosoma cruzi (POPTc80) is associated with the entry of trypomastigotes into mammalian host cells. In this study, the gene coding to prolyl oligopeptidase of Trypanosoma brucei (POPTb) was identified and characterized. It is represented by a single copy per haploid genome of the parasite and its deduced amino acid sequence shares 77% identity with POPTc80. Secondary structure predictions demonstrated that POPTb shows the highly preserved secondary structure composition and arrangement of prolyl oligopeptidases. Active recombinant POP Tb, produced in E. coli , displayed enzymatic activity on peptides containing Pro at P1 position and collagen at a slightly alkaline pH. Its enzymatic activity was highly sensitive to POPTc80 inhibitors. Furthermore, these inhibitors arrested growth of procyclic and bloodstream T. brucei forms in a dose-dependent manner. These data suggest that POPTb activity is required for normal parasite development. |
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