Defensins are basic, cysteine-rich antimicrobial peptides that are important components of plant defense against pathogens. Previously, we isolated a defensin, PvD1, from Phaseolus vulgaris L. (common bean) seeds.

The Aedes aegypti mosquito is responsible for transmitting pathogens such as the Dengue, Zika, and Chikungunya viruses. The peritrophic matrix (PM) is an extracellular chitin-rich structure that lines the midgut of arthropods, providing a crucial protective barrier for the gut epithelium against mechanical damage, ingested pathogens, and toxic substances. During blood digestion, hemoglobin is lysed, releasing free heme into the midgut lumen. Part of this heme binds strongly to the PM, mitigating its harmful effects on the mosquito epithelial cells. Our study focused on investigating the interaction dynamic between hemoglobin and the PM during blood digestion in A. aegypti. Optical microscopy was employed to observe the temporal progression of blood digestion in the A. aegypti midgut, highlighting significant morphological changes in the blood bolus. An electrophoresis analysis revealed distinct protein bands in the PM extract, some of which were associated with hemoglobin and its subunits. The presence of PM-associated hemoglobin was confirmed by amino-terminal sequencing and an immunoblot analysis using anti-hemoglobin antibodies. Furthermore, fluorescence microscopy revealed overlapping labeling between hemoglobin and chitin, suggesting an interaction between hemoglobin and PM chitin. Corroborating these results, hemoglobin showed an affinity with chitin in the chromatography and molecular docking assays, in which the hemoglobin subunits interacted with the oligosaccharide (NAG)₄. Thus, hemoglobin may perform a function similar to that of peritrophins. Further experiments demonstrated the protective role of the PM against hemoglobin proteolysis during blood digestion. Overall, this study provides valuable insights into the intricate interactions between hemoglobin and the PM, enhancing our understanding of mosquito digestive physiology and potentially contributing to the development of vector control strategies.

Viperine and crotaline snake venoms contain one or more hemorrhagic metalloproteinases called hemorrhagins. The most potent hemorrhagins belong to P-III class and have, in adition to the protease domain, a disintegrin-like and a cysteine-rich domains. Although proteolytic degradation of vascular endothelium basement membrane has been established to be the main factor responsible for hemorrhage, several studies reveal other factors that actually do facilitate this process. In this study we report the identification of an P-I class hemorrhagin from Bothrops atrox venom. We have formerly purified an P-III class hemorrhagin from the same venom. Although exhibiting a higher proteolytic activity, the P-I class hemorrhagin showed to be a less efficient hemorrhagin when compared in vivo with the previously described P-III class metalloprotease.

Ric c1, an allergenic protein from castor oil plants (Ricinus communis), is an insect α-amylase inhibitor that has become an occupational allergen. Ric c1 can cross-react with allergens from wheat, soybean, peanut, shrimp, fish, gluten, house dust, tobacco and air fungus, thereby amplifying the concern and risks caused by castor oil plants (COP) allergens. Two continuous IgE-binding epitopes were identified in Ric c1, both containing glutamic acid residues involved in IgE-binding and allergic challenges. We produced recombinant Ric c1 (rRic c1) in Escherichia coli, using primers from foliar castor oil plant DNA, and a mutant (Glu-Leu) recombinant protein (mrRic c1) in the same system using synthetic genes. rRic c1 preserved both allergenic and α-amylase inhibitory properties, and mrRic c1 drastically reduced allergenic properties. These results can help to establish meaningful relationships between structure, defence and allergenicity, important steps for producing engineered plants and developing new approaches for immunotherapy.