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Mass spectrometry-based identification of allergens from Curvularia pallescens, a prevalent aerospore in India Proteins and proteomics
- Dey, Debarati, Saha, Bodhisattwa, Sircar, Gaurab, Ghosal, Kavita, Bhattacharya, Swati Gupta
- Biochimica et biophysica acta 2016 v.1864 no.7 pp. 869-879
- Curvularia pallescens, allergens, databases, fructose-bisphosphate aldolase, fungi, gels, histamine, human health, hypersensitivity, immunoglobulin E, mass spectrometry, patients, proteinases, proteome, proteomics, therapeutics, vacuoles, India
- The worldwide prevalence of fungal allergy in recent years has augmented mining allergens from yet unexplored ones. Curvularia pallescens (CP) being a dominant aerospore in India and a major sensitiser on a wide range of allergic population, pose a serious threat to human health. Therefore, we aimed to identify novel allergens from CP in our present study. A cohort of 22 CP-sensitised patients was selected by positive Skin prick grade. Individual sera exhibited elevated specific IgE level and significant histamine release on a challenge with antigenic extract of CP. First gel-based profiling of CP proteome was done by 1- and 2-dimensional gel. Parallel 1- and 2-dimensional immunoblot were performed applying individual as well as pooled patient sera. Identification of the sero-reactive spots from the 2-dimensional gel was found to be challenging as CP was not previously sequenced. Hence, mass spectrometry-based proteomic workflow consisting of conventional database search was not alone sufficient. Therefore, de novo sequencing preceded homology search was implemented for further identification. Altogether 11 allergenic proteins including Brn-1, vacuolar protease, and fructose-bis-phosphate aldolase were identified with high statistical confidence (p<0.05). This is the first study to report on any allergens from CP. This kind of proteome-based analysis provided a catalogue of CP allergens that would lead an improved way of diagnosis and therapy of CP-related allergy.