RESUMEN
The hazelnut allergen Cor a 14 belongs to the 2S albumins, a family of heterodimeric seed storage proteins exhibiting a high degree of structural diversity. Given its relevance as an allergen and the potential to elicit severe reactions, elucidation of the sequence heterogeneity of naturally occurring Cor a 14 is essential for the development of reliable diagnostics and risk evaluation. We therefore performed a comprehensive survey on the proteoforms of Cor a 14 and determined their quantitative distribution in three different hazelnut cultivars by a combinatory HPLC-HRMS approach including bottom-up and intact mass analysis. Compared with the Cor a 14 prototype sequence, we identified three sequence polymorphisms, two of the small and one of the large subunit, and elucidated their specific pairing on the protein level. Furthermore, we located a pronounced microheterogeneity on the protein termini and, for the first time, provide data on varying proteoform patterns between different cultivars of an allergenic seed. Together, these data present the basis for a more detailed investigation on the allergenicity of Cor a 14 in different cultivars and constitute, to be best of our knowledge, the largest set of proteoforms so far reported for a 2S albumin.
Asunto(s)
Alérgenos/química , Antígenos de Plantas/química , Polimorfismo de Nucleótido Simple , Subunidades de Proteína/química , Proteínas de Almacenamiento de Semillas/química , Alérgenos/genética , Alérgenos/aislamiento & purificación , Secuencia de Aminoácidos , Antígenos de Plantas/genética , Antígenos de Plantas/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Corylus/química , Expresión Génica , Humanos , Espectrometría de Masas , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/aislamiento & purificación , Subunidades de Proteína/genética , Subunidades de Proteína/aislamiento & purificación , Proteínas de Almacenamiento de Semillas/genética , Proteínas de Almacenamiento de Semillas/aislamiento & purificaciónRESUMEN
Background: Omics technologies have been widely applied in different fields, among which, proteomics is gaining increasing interest for its application to the authenticity of food products. MS, typically coupled with LC, represents a key technique for proteomics-related studies dedicated to fish and other seafood products by using a bottom-up approach. Objective and Methods: In this paper, the optimization of an untargeted proteomics-based method using LC separation and MS detection relying on a quadrupole time-of-flight mass spectrometer is described and applied to the analysis of Canadian farmed and wild-type salmon, followed by statistical analysis based on principal component (PC) analysis. Results and Conclusions: This untargeted approach, using a data-independent acquisition MS scheme, demonstrated the ability to effectively discriminate salmon belonging to the two classes. Furthermore, selected peptides showing high loadings on PC1 could represent potential candidate peptide markers able to discriminate farmed from wild-type salmon samples in the future.