Publication: Effects of Ser47-point mutation on conformation structure and allergenicity of the allergen of Der p 2, a major house dust mite allergen
Issued Date
2019-01-01
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20927363
20927355
20927355
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2-s2.0-85057976764
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Mahidol University
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SCOPUS
Bibliographic Citation
Allergy, Asthma and Immunology Research. Vol.11, No.1 (2019), 129-142
Suggested Citation
Bhakkawarat Kulwanich, Sasipa Thanyaratsrisakul, Orathai Jirapongsananuruk, Belinda J. Hales, Wayne R. Thomas, Surapon Piboonpocanun Effects of Ser47-point mutation on conformation structure and allergenicity of the allergen of Der p 2, a major house dust mite allergen. Allergy, Asthma and Immunology Research. Vol.11, No.1 (2019), 129-142. doi:10.4168/aair.2019.11.1.129 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/51139
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Title
Effects of Ser47-point mutation on conformation structure and allergenicity of the allergen of Der p 2, a major house dust mite allergen
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Abstract
Copyright © 2019 The Korean Academy of Asthma, Allergy and Clinical Immunology • The Korean Academy of Pediatric Allergy and Respiratory Disease. Purpose: Hypoallergenic recombinant Der p 2 has been produced by various genetic manipulations, but mutation of a naturally polymorphic amino acid residue known to affect IgE binding has not been studied. This study aimed to determine the effect of a point mutation (S47W) of residue 47 of Der p 2 on its structure and immunoglobulin (Ig) E binding. Its ability to induce pro-inflammatory responses and to induce blocking IgG antibody was also determined. Methods: S47 of recombinant Der p 2.0110, one of the predominant variants in Bangkok, was mutated to W (S47W). S47W secreted from Pichia pastoris was examined for secondary structure and for the formation of a hydrophobic cavity by 8-Anilino-1-naphthalenesulfonic acid (ANS) staining. Monoclonal and human IgE-antibody binding was determined by enzyme-linked immunosorbent assay. Allergen-induced degranulation by human epsilon receptor expressed-rat basophil was determined. Stimulation of the pro-inflammatory cytokine interleukin (IL)-8 release from human bronchial epithelial (BEAS2B) cells and inhibition of IgE binding to the wild type allergen by S47W-induced IgG were determined. Results: S47W reduced secondary structure and failed to bind the hydrophobic ANS ligand as well as a monoclonal antibody known to be dependent on the nature of the side chain of residue 114 in an adjacent loop. It could also not stimulate IL-8 release from BEAS2B cells. IgE from house dust mite (HDM)-allergic Thais bound S47W with 100-fold weaker avidity, whereas IgE of HDM-allergic Australians did not. S47W still induced basophil degranulation, although requiring higher concentrations for some subjects. Anti-S47W antiserum-immunized mice blocked the binding of human IgE to wild type Der p 2. Conclusions: The mutant S47W had altered structure and reduced ability to stimulate pro-inflammatory responses and to bind IgE, but retained its ability to induce blocking antibodies. It thus represents a hypoallergen produced by a single mutation of a non-solvent-accessible amino acid.