Isolation and biochemical characterization of a thaumatin-like kiwi allergen
Само за регистроване кориснике
2002
Аутори
Gavrović-Jankulović, MarijaĆirković-Veličković, Tanja
Vučković, O.
Atanasković-Marković, Marina
Petersen, A.
Gojgić-Cvijović, Gordana D.
Burazer, Lidija M.
Jankov, Ratko M.
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Background: Kiwi fruit allergy, as well as its association with hypersensitivity to other foods and to pollen, has been extensively reported in the last few years. Several IgE-binding components have been detected in kiwi extract, but only one 30-kd allergen has been isolated; it was identified as actinidin (Act c 1). Recently, we have reported a 24-kd kiwi protein to be a potential major allergen in a group of patients with oral allergy syndrome (OAS). Objective: The aim of this study was to purify and characterize the 24-kd kiwi allergen biochemically. Methods: Seven polysensitized patients with OAS to kiwi were used in this study. The kiwi allergen was isolated by using a combination of gel permeation, ion exchange, and immobilized metal ion affinity chromatography. Its biochemical characterization included determination of its isoelectric point, molecular weight, N-terminal sequencing, concanavalin A-binding ability, digestibility in simulated gastric fluid, and antifungal activity.... Western blotting, 2-dimensional PAGE immunoblotting, and skin prick tests were performed to characterize the isolated protein immunochemically. Results: All 7 patients recognized the isolated 24-kd kiwi protein as an allergen. The isolated protein consisted of 2 isoforms with isoelectric points of 9.4 and 9.5 migrated as one protein band of 20 kd after SDS-PAGE under nonreducing conditions or at 24 kd under reducing conditions. The partial N-terminal sequence revealed that it is a thaumatin-like protein (TLP) with concanavalin A-binding ability. The protein showed antifungal activity toward Saccharomyces carlsbergensis, and Candida albicans. The protein was degraded by the simulated gastric fluid within 1 minute. Both isoforms bound IgE from a pool of sera in a 2-dimensional PAGE inummoblot. The TLP elicited positive skin prick test responses in 4 (80%) of 5 patients with OAS. Conclusion: This study reported isolation and full characterization of a new kiwi allergen, TLP (isoelectric points of 9.4 and 9.5 and molecular weight of 24 kd), which belongs to the family of pathogenesis-related proteins. The isolated protein expressed antifungal activity toward S carlsbergensis and C albicans.
Кључне речи:
allergen purification / antifungal activity / food allergen / kiwi fruit / pathogenesis-related protein / thaumatin-like proteinИзвор:
Journal of Allergy and Clinical Immunology, 2002, 110, 5, 805-810Издавач:
- Mosby, Inc, St Louis
DOI: 10.1067/mai.2002.128947
ISSN: 0091-6749
PubMed: 12417892
WoS: 000179082500018
Scopus: 2-s2.0-0036858690
Колекције
Институција/група
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Gavrović-Jankulović, Marija AU - Ćirković-Veličković, Tanja AU - Vučković, O. AU - Atanasković-Marković, Marina AU - Petersen, A. AU - Gojgić-Cvijović, Gordana D. AU - Burazer, Lidija M. AU - Jankov, Ratko M. PY - 2002 UR - https://cherry.chem.bg.ac.rs/handle/123456789/513 AB - Background: Kiwi fruit allergy, as well as its association with hypersensitivity to other foods and to pollen, has been extensively reported in the last few years. Several IgE-binding components have been detected in kiwi extract, but only one 30-kd allergen has been isolated; it was identified as actinidin (Act c 1). Recently, we have reported a 24-kd kiwi protein to be a potential major allergen in a group of patients with oral allergy syndrome (OAS). Objective: The aim of this study was to purify and characterize the 24-kd kiwi allergen biochemically. Methods: Seven polysensitized patients with OAS to kiwi were used in this study. The kiwi allergen was isolated by using a combination of gel permeation, ion exchange, and immobilized metal ion affinity chromatography. Its biochemical characterization included determination of its isoelectric point, molecular weight, N-terminal sequencing, concanavalin A-binding ability, digestibility in simulated gastric fluid, and antifungal activity. Western blotting, 2-dimensional PAGE immunoblotting, and skin prick tests were performed to characterize the isolated protein immunochemically. Results: All 7 patients recognized the isolated 24-kd kiwi protein as an allergen. The isolated protein consisted of 2 isoforms with isoelectric points of 9.4 and 9.5 migrated as one protein band of 20 kd after SDS-PAGE under nonreducing conditions or at 24 kd under reducing conditions. The partial N-terminal sequence revealed that it is a thaumatin-like protein (TLP) with concanavalin A-binding ability. The protein showed antifungal activity toward Saccharomyces carlsbergensis, and Candida albicans. The protein was degraded by the simulated gastric fluid within 1 minute. Both isoforms bound IgE from a pool of sera in a 2-dimensional PAGE inummoblot. The TLP elicited positive skin prick test responses in 4 (80%) of 5 patients with OAS. Conclusion: This study reported isolation and full characterization of a new kiwi allergen, TLP (isoelectric points of 9.4 and 9.5 and molecular weight of 24 kd), which belongs to the family of pathogenesis-related proteins. The isolated protein expressed antifungal activity toward S carlsbergensis and C albicans. PB - Mosby, Inc, St Louis T2 - Journal of Allergy and Clinical Immunology T1 - Isolation and biochemical characterization of a thaumatin-like kiwi allergen VL - 110 IS - 5 SP - 805 EP - 810 DO - 10.1067/mai.2002.128947 ER -
@article{ author = "Gavrović-Jankulović, Marija and Ćirković-Veličković, Tanja and Vučković, O. and Atanasković-Marković, Marina and Petersen, A. and Gojgić-Cvijović, Gordana D. and Burazer, Lidija M. and Jankov, Ratko M.", year = "2002", abstract = "Background: Kiwi fruit allergy, as well as its association with hypersensitivity to other foods and to pollen, has been extensively reported in the last few years. Several IgE-binding components have been detected in kiwi extract, but only one 30-kd allergen has been isolated; it was identified as actinidin (Act c 1). Recently, we have reported a 24-kd kiwi protein to be a potential major allergen in a group of patients with oral allergy syndrome (OAS). Objective: The aim of this study was to purify and characterize the 24-kd kiwi allergen biochemically. Methods: Seven polysensitized patients with OAS to kiwi were used in this study. The kiwi allergen was isolated by using a combination of gel permeation, ion exchange, and immobilized metal ion affinity chromatography. Its biochemical characterization included determination of its isoelectric point, molecular weight, N-terminal sequencing, concanavalin A-binding ability, digestibility in simulated gastric fluid, and antifungal activity. Western blotting, 2-dimensional PAGE immunoblotting, and skin prick tests were performed to characterize the isolated protein immunochemically. Results: All 7 patients recognized the isolated 24-kd kiwi protein as an allergen. The isolated protein consisted of 2 isoforms with isoelectric points of 9.4 and 9.5 migrated as one protein band of 20 kd after SDS-PAGE under nonreducing conditions or at 24 kd under reducing conditions. The partial N-terminal sequence revealed that it is a thaumatin-like protein (TLP) with concanavalin A-binding ability. The protein showed antifungal activity toward Saccharomyces carlsbergensis, and Candida albicans. The protein was degraded by the simulated gastric fluid within 1 minute. Both isoforms bound IgE from a pool of sera in a 2-dimensional PAGE inummoblot. The TLP elicited positive skin prick test responses in 4 (80%) of 5 patients with OAS. Conclusion: This study reported isolation and full characterization of a new kiwi allergen, TLP (isoelectric points of 9.4 and 9.5 and molecular weight of 24 kd), which belongs to the family of pathogenesis-related proteins. The isolated protein expressed antifungal activity toward S carlsbergensis and C albicans.", publisher = "Mosby, Inc, St Louis", journal = "Journal of Allergy and Clinical Immunology", title = "Isolation and biochemical characterization of a thaumatin-like kiwi allergen", volume = "110", number = "5", pages = "805-810", doi = "10.1067/mai.2002.128947" }
Gavrović-Jankulović, M., Ćirković-Veličković, T., Vučković, O., Atanasković-Marković, M., Petersen, A., Gojgić-Cvijović, G. D., Burazer, L. M.,& Jankov, R. M.. (2002). Isolation and biochemical characterization of a thaumatin-like kiwi allergen. in Journal of Allergy and Clinical Immunology Mosby, Inc, St Louis., 110(5), 805-810. https://doi.org/10.1067/mai.2002.128947
Gavrović-Jankulović M, Ćirković-Veličković T, Vučković O, Atanasković-Marković M, Petersen A, Gojgić-Cvijović GD, Burazer LM, Jankov RM. Isolation and biochemical characterization of a thaumatin-like kiwi allergen. in Journal of Allergy and Clinical Immunology. 2002;110(5):805-810. doi:10.1067/mai.2002.128947 .
Gavrović-Jankulović, Marija, Ćirković-Veličković, Tanja, Vučković, O., Atanasković-Marković, Marina, Petersen, A., Gojgić-Cvijović, Gordana D., Burazer, Lidija M., Jankov, Ratko M., "Isolation and biochemical characterization of a thaumatin-like kiwi allergen" in Journal of Allergy and Clinical Immunology, 110, no. 5 (2002):805-810, https://doi.org/10.1067/mai.2002.128947 . .