Pollen from different tree and grass species represent the greatest inducers of type I allergy worldwide. Oriental plane trees, as Platanus orientalis, are an important source of airborne allergens in cities of the southwest Asia and southeast Europe. This study was aimed to identify relevant allergens of P. orientalis pollen and to ascertain whether P. orientalis allergens have cross-reactivity with related plane trees, such as Platanus acerifolia and Platanus occidentalis pollen components. Nineteen patients with a clinical history of reaction to P. orientalis pollen and a positive skin-prick test (SPT) to P. orientalis pollen extract were included in this study. Identification of IgE-binding proteins in Platanus pollen extracts was elucidated by immunoblotting using sera from P. orientalis pollen-sensitive patients. Cross-reactivity studies among P. orientalis allergens and relevant species was evaluated by immunoblot-inhibition and ELISA inhibition assays. All the patients were polysensitive and exhibited positive SPT to grass and tree pollen allergens. The IgE-binding pattern of P. orientalis pollen extract was well defined. The inhibition experiments showed that the capacity of P. occidentalis was greater than P. acerifolia pollen extract in preventing the reactivity of immobilized P. orientalis pollen extract with the IgE antibodies of patients. The 28-kDa molecule was the most frequent allergen among nine IgE-binding proteins of P. orientalis pollen. The IgE-reactive components of P. orientalis pollen showed a higher level of cross-reactivity with P. occidentalis pollen in comparison with P. acerifolia pollen.
SummaryBackground: Profilins are highly cross-reactive allergens which bind IgE antibodies of almost 20% of plant-allergic patients. This study is aimed at investigating cross-reactivity of melon profilin with other plant profilins and the role of the linear and conformational epitopes in human IgE cross-reactivity.
A simple one-step method for the purification of recombinant His-tagged profilin from the bacterial cell lysate is reported. Noting the greater ease with which hexahistidine-tagged proteins can be metalprecipitated compared with unwanted protein impurities, we investigated the effect of lysis-buffer additives and optimization of other conditions to recover selectively desired proteins in a one-step metal precipitation without using biopolymers. Purification of the His-tagged melon (Cucumis melo) profilin was used to demonstrate the utility of this method and up to 80% recovery with a purity of 98% was achieved. This method obtained a yield of the protein nearly comparable with that obtained using metal-affinity chromatography. This purification procedure can reduce the time and cost of the purification process, especially on a large scale.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.