In-depth exploration of Hevea brasiliensis latex proteome and “hidden allergens” via combinatorial peptide ligand libraries

Division of Genetics and Cell Biology, San Raffaele Scientific Institute, 20132 Milano, Italy.
Journal of proteomics (Impact Factor: 3.89). 03/2010; 73(7):1368-80. DOI: 10.1016/j.jprot.2010.03.002
Source: PubMed


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.

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    • "A 33 kDa rice allergen was identified as a glyoxalase [37]. A proteasome subunit protein was already reported as a Hevea brasiliensis latex allergen [38]. Finally, as general stress protein 39, several fungal allergens belong to SDR protein family [35-40]. "
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    • "In order to eliminate pigments and other polymeric materials [12] 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). "

    Revue Française d'Allergologie; 04/2012
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    • "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. [12]. 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. "
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