ABSTRACT
The description of Plasmodium ookinete surface proteins and their participation in the complex process of mosquito midgut invasion is still incomplete. In this study, using phage display, a consensus peptide sequence (PWWP) was identified in phages that bound to the Plasmodium berghei ookinete surface and, in selected phages, bound to actin and enolase in overlay assays with ookinete protein extracts. Actin was localized on the surface of fresh live ookinetes by immunofluorescence and electron microscopy using specific antibodies. The overall results indicated that enolase and actin can be located on the surface of ookinetes, and suggest that they could participate in Plasmodium invasion of the mosquito midgut.
Subject(s)
Actins/metabolism , Aedes/metabolism , Anopheles/metabolism , Digestive System/metabolism , Life Cycle Stages/genetics , Peptides/metabolism , Phosphopyruvate Hydratase/metabolism , Plasmodium berghei , Aedes/genetics , Aedes/parasitology , Amino Acid Sequence , Animals , Anopheles/genetics , Anopheles/parasitology , Cell Extracts , Cells, Cultured , Conserved Sequence , Digestive System/parasitology , Fluorescent Antibody Technique , Malaria/metabolism , Malaria/parasitology , Male , Mice , Mice, Inbred BALB C , Microscopy, Electron , Molecular Sequence Data , Peptide Library , Peptides/chemistry , Peptides/genetics , Plasmodium berghei/genetics , Plasmodium berghei/metabolism , Protozoan Proteins/metabolism , Rodent Diseases/metabolism , Rodent Diseases/parasitologyABSTRACT
Salivary glands of female mosquitoes produce proteins, not completely described yet, that participate in carbohydrate and blood feeding. Here, we report an acidic glycoprotein of 35 kDa (GP35 ANOAL) secreted in the saliva of the malaria vector mosquito Anopheles albimanus. GP35 ANOAL is produced exclusively in the distal lateral lobes of adult female salivary glands, it has a pI of 4.45 and is negatively stained by regular silver stain. An 888 bp cDNA clone encoding a predicted product of 240 amino acids has a signal peptide, potential post-translational modification sites, and a disintegrin signature RGD. The GP35 ANOAL sequence depicts high similarities with the 30 kDa saliva allergen of Aedes aegypti, 30 kDa allergen-like hypothetical proteins, and GE-rich proteins present in several Anopheles species, as well as in Ae. albopictus and Culex pipiens quinquefasciatus. The function of this protein family is still unknown.
Subject(s)
Anopheles/metabolism , Glycoproteins/metabolism , Insect Proteins/metabolism , Amino Acid Sequence , Animals , Anopheles/genetics , Anopheles/growth & development , Base Sequence , Female , Glycoproteins/genetics , Insect Proteins/genetics , Molecular Sequence Data , RNA, Messenger/metabolism , Salivary Glands/metabolismABSTRACT
Malaria is one of the most important parasitic diseases in the world and a major public health problem because of emerging drug-resistant strains of Plasmodium. A number of synthetic and natural compounds are now being analysed to develop more effective antimalarial drugs. We investigated the effect of homeopathic preparations of Eupatorium perfoliatum and Arsenicum album on parasitemia using a rodent malaria model. We found significant inhibitory effect on parasite multiplication with both medications with a level of 60% for Eupatorium perfoliatum at a 30 CH potency. Arsenicum album 0/6 gave 70% inhibition but this was less stable than Eupatorium perfoliatum. The number of schizonts was higher in animals treated with homeopathic medications. Although the mechanism of action is unknown, these agents would be good candidates as alternative or complementary medications in the treatment of malaria.