Class I chitinases, the panallergens responsible for the latex-fruit syndrome, are induced by ethylene treatment and inactivated by heating Rosa Sánchez-Monge,

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Presentation transcript:

Class I chitinases, the panallergens responsible for the latex-fruit syndrome, are induced by ethylene treatment and inactivated by heating Rosa Sánchez-Monge, PhDa, Carlos Blanco, PhDb, Araceli Díaz Perales, BSa, Carmen Collada, PhDc, Teresa Carrillo, PhDb, Cipriano Aragoncillo, PhDc, Gabriel Salcedo, PhDa Journal of Allergy and Clinical Immunology Volume 106, Issue 1, Pages 190-195 (July 2000) DOI: 10.1067/mai.2000.107599 Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 1 SDS-PAGE separation of crude extracts (15 μg) from avocado (lane 1) ; untreated green beans (lane 2) ; 24-hour (lane 3) , 48-hour (lane 4) , and 72-hour (lane 5) ethylene-treated green beans; heat-treated (lane 7) green beans; and untreated white beans (lane 6) . Replica gels were stained with Coomassie blue (A) or electrotransferred to PVDF membranes and immunoblotted with monospecific polyclonal antibodies to plant chitinases (B) or a serum pool from patients with latex-fruit allergy (C) . Journal of Allergy and Clinical Immunology 2000 106, 190-195DOI: (10.1067/mai.2000.107599) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 2 SDS-PAGE separation of crude extracts from untreated green beans (lane 1) , 72-hour ethylene-treated green beans (lane 2) , white beans (lane 4) , and 32-kd purified proteins from ethylene-treated green beans (lane 3) and white beans (lane 5) . Replica gels were stained with Coomassie blue (A) or electrotransferred to PVDF membranes and immunodetected with monospecific polyclonal antibodies to plant chitinases (B) or a serum pool from patients with latex-fruit allergy (C) . Journal of Allergy and Clinical Immunology 2000 106, 190-195DOI: (10.1067/mai.2000.107599) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 3 N-terminal amino acid sequences of green bean class I chitinase PvChI and white bean phytohemagglutinin PvPhy aligned with that previously reported for P vulgaris PHA-L.25 X , Undetermined residue; PHA, phytohemagglutinin. Journal of Allergy and Clinical Immunology 2000 106, 190-195DOI: (10.1067/mai.2000.107599) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 4 Immunoblot inhibition assays with the purified class I chitinases Prs a 1 and PvChI as inhibitors. PBS extracts (15 μg) from avocado (lanes 1) or 72-hour ethylene-treated green beans (lanes 2) were fractionated by means of SDS-PAGE and electrotransferred to PVDF membranes. Immunodetection was performed with a pool of sera from patients with latex-fruit allergy preincubated with BSA (+BSA ; negative control) or with the major avocado allergen (+Prs a 1) and green bean chitinase (+PvChI) . Five micrograms of each inhibitor was tested. Journal of Allergy and Clinical Immunology 2000 106, 190-195DOI: (10.1067/mai.2000.107599) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 5 SDS-PAGE separation of untreated (–) and heat-treated (+) purified green bean class I chitinase (PvChI), peach lipid-transfer protein (LTP) , BSA, and an avocado protein preparation enriched in chitinases (AvF) . Gels were stained with Coomassie blue (A and D ) or electrotransferred to PVDF membranes and immunoblotted with monospecific polyclonal antibodies to plant chitinases (B) or a serum pool from patients with latex-fruit allergy (C) . Journal of Allergy and Clinical Immunology 2000 106, 190-195DOI: (10.1067/mai.2000.107599) Copyright © 2000 Mosby, Inc. Terms and Conditions