The proteome of Hevea brasiliensis latex has been explored in depth via combinatorial peptide ligand libraries. A total of 300 unique gene products have been identified in this latex, whose proteome has been largely unknown up to the present. In search for unknown allergens, control latex and eluates from the ligand libraries have been fractionated by two-dimensional mapping, blotted and confronted with sera of 18 patients. In addition to the already known and named Hevea major allergens, we have unambiguously detected several others like, for instance: heat shock protein (81 kDa), proteasome subunit (30 kDa), protease inhibitor (8 kDa), hevamine A (43 kDa) and glyceraldehyde-3-phosphate dehydrogenase (37 kDa). Gene Ontology analysis of analyzed fractions has shown that major functions are substantially unchanged after sample treatment, while novel biological functions appeared that were undetectable in the crude sample.
"A 33 kDa rice allergen was identified as a glyoxalase . A proteasome subunit protein was already reported as a Hevea brasiliensis latex allergen . Finally, as general stress protein 39, several fungal allergens belong to SDR protein family [35-40]. "
[Show abstract][Hide abstract] ABSTRACT: Allergies are nearly always triggered by protein molecules and the majority of individuals with documented immunologic reactions to foods exhibit IgE hypersensitivity reactions. In this study we aimed to understand if natural differences, at proteomic level, between maize populations, may induce different IgE binding proteins profiles among maize-allergic individuals. We also intended to deepen our knowledge on maize IgE binding proteins.
In order to accomplish this goal we have used proteomic tools (SDS-PAGE and 2-D gel electrophoresis followed by western blot) and tested plasma IgE reactivity from four maize-allergic individuals against four different protein fractions (albumins, globulins, glutelins and prolamins) of three different maize cultivars. We have observed that maize cultivars have different proteomes that result in different IgE binding proteins profiles when tested against plasma from maize-allergic individuals. We could identify 19 different maize IgE binding proteins, 11 of which were unknown to date. Moreover, we found that most (89.5%) of the 19 identified potential maize allergens could be related to plant stress.
These results lead us to conclude that, within each species, plant allergenic potential varies with genotype. Moreover, considering the stress-related IgE binding proteins identified, we hypothesise that the environment, particularly stress conditions, may alter IgE binding protein profiles of plant components.
"In order to eliminate pigments and other polymeric materials  both PUN and PBS extracts were precipitated by addition of ammonium sulfate up to 90% saturation or, respectively, 66 g (for 100 mL) and 82.5 g (for 125 mL). These two mixtures were gently agitated at 4 °C overnight and then the supernatants eliminated by centrifugation (30 min at 18 000 g at 4 °C). "
"In contrast, we provide a comprehensive MS analysis covering all three latex subproteomes of T. brevicorniculatum. Without the enrichment of low-abundance proteins, our method was sensitive enough to detect scarce proteins such as annexin D1 (Q9SYT0), proteasome subunit alpha type-4 (O82530), and putative DNA repair protein RAD23-4 (Q84L39) as reported only after the enrichment of low-abundance proteins by D´Amato et al. . These results confirm that we have developed robust and sensitive methods for investigating the T. brevicorniculatum latex proteome while simultaneously providing a detailed overview of abundant latex proteins. "
[Show abstract][Hide abstract] ABSTRACT: Many plants produce latex, a specialized, metabolically active cytoplasm. This is generally regarded as a defensive trait but latex may also possess additional functions. We investigated the role of latex in the dandelion species Taraxacum brevicorniculatum that contains considerable amounts of high-quality natural rubber by carrying out a comprehensive analysis of the latex proteome. We developed reliable protocols for the preparation of protein samples for one-dimensional gel electrophoresis, two-dimensional gel electrophoresis, and subsequent mass spectrometry analysis, which led to 278 unique identifications. A gene ontology classification system based on comparisons with known Arabidopsis thaliana root proteins showed that dandelion proteins involved in lipid metabolism and transport were enriched in the latex proteome, whereas those involved in stress responses were not. We also found that proteins involved in rubber biosynthesis were distributed among different fractions of the latex proteome.
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