Rational Design, Structure-Activity Relationship, and Immunogenicity of Hypoallergenic Pru p 3 Variants.
Mol Nutr Food Res
; 63(18): e1900336, 2019 09.
Article
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| MEDLINE
| ID: mdl-31207117
ABSTRACT
SCOPE Allergies to lipid transfer proteins involve severe adverse reactions; thus, effective and sustainable therapies are desired. Previous attempts disrupting disulfide bonds failed to maintain immunogenicity; thus, the aim is to design novel hypoallergenic Pru p 3 variants and evaluate the applicability for treatment of peach allergy. METHODS AND RESULTS:
Pru p 3 proline variant (PV) designed using in silico mutagenesis, cysteine variant (CV), and wild-type Pru p 3 (WT) are purified from Escherichia coli. Variants display homogenous and stable protein conformations with an altered secondary structure in circular dichroism. PV shows enhanced long-term storage capacities compared to CV similar to the highly stable WT. Using sera of 33 peach allergic patients, IgE-binding activity is reduced by 97% (PV) and 71% (CV) compared to WT. Both molecules show strong hypoallergenicity in Pru p 3 ImmunoCAP cross-inhibition and histamine release assays. Immunogenicity of PV is demonstrated with a phosphate-based adjuvant formulation in a mouse model.CONCLUSIONS:
An in silico approach is used to generate a PV without targeting disulfide bonds, T cell epitopes, or previously reported IgE epitopes of Pru p 3. PV is strongly hypoallergenic while structurally stable and immunogenic, thus representing a promising candidate for peach allergen immunotherapy.Palabras clave
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Base de datos:
MEDLINE
Asunto principal:
Proteínas de Plantas
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Proteínas Recombinantes
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Antígenos de Plantas
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Hipersensibilidad a los Alimentos
Idioma:
En
Revista:
Mol Nutr Food Res
Asunto de la revista:
CIENCIAS DA NUTRICAO
Año:
2019
Tipo del documento:
Article