title: Mass spectrometry-based identification of allergens from Curvularia pallescens, a prevalent aerospore in India.
abstract: 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.
authors.affiliation: Division of Plant Biology, Bose Institute, 93/1, Acharya Prafulla Chandra Road, Kolkata-700009, West Bengal, India. Electronic address: debaratidey03@gmail.com.
authors.forename: Debarati
authors.initials: D
authors.lastname: Dey
authors.affiliation: Division of Plant Biology, Bose Institute, 93/1, Acharya Prafulla Chandra Road, Kolkata-700009, West Bengal, India. Electronic address: bodhisattwasaha@gmail.com.
authors.forename: Bodhisattwa
authors.initials: B
authors.lastname: Saha
authors.affiliation: Division of Plant Biology, Bose Institute, 93/1, Acharya Prafulla Chandra Road, Kolkata-700009, West Bengal, India. Electronic address: sircar.gaurab129@gmail.com.
authors.forename: Gaurab
authors.initials: G
authors.lastname: Sircar
authors.affiliation: Division of Plant Biology, Bose Institute, 93/1, Acharya Prafulla Chandra Road, Kolkata-700009, West Bengal, India. Electronic address: kavita.ghosal@gmail.com.
authors.forename: Kavita
authors.initials: K
authors.lastname: Ghosal
authors.affiliation: Division of Plant Biology, Bose Institute, 93/1, Acharya Prafulla Chandra Road, Kolkata-700009, West Bengal, India. Electronic address: swati@jcbose.ac.in.
authors.forename: Swati Gupta
authors.initials: SG
authors.lastname: Bhattacharya
mesh_terms: D000485:Allergens / Q000032:analysis*; D001203:Ascomycota / Q000276:immunology*; D006801:Humans; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013172:Spores, Fungal
chemical_list: D000485:Allergens
keywords: Allergen; Curvularia pallescens; Fungal allergic patients; Immunoblot; MALDI TOF/TOF