title: Shotgun proteome analysis of beer and the immunogenic potential of beer polypeptides.
abstract: The majority of beer proteins originate from barley (Hordeum vulgare) which is used for brewing. Barley is known to contain celiacogenic gliadin-like prolamins (hordeins) along with other immunogenic proteins which endure malt proteases and the harsh conditions of brewing. In addition, a multitude of peptides that may retain or even amplify the immune-stimulating potential is released in beer because of proteolysis. The comprehensive annotation of the beer proteome is challenged both by the high concentration range of the protein entities and by a severe degree of processing-induced modifications. Overcoming the pitfalls of the classical two-dimensional electrophoresis approach coupled to mass spectrometry (MS), the gel-free shotgun proteomic analysis expanded the current inventory of a popular Italian beer to 33 gene products, including traces of intact B- and D-hordeins and 10 proteins from Saccharomyces spp. The high performance liquid chromatography-electrospray MS/MS peptidomic analysis of the low-molecular weight beer components disclosed a panel of hordein-derived peptides that encrypt gluten-like sequence motifs, potentially harmful to celiacs. The presence of antigliadin IgA-immunoresponsive prolamins was assayed by Western and dot blot using sera of N=4 celiac patients. Gliadin-reactive T-cell lines isolated from the intestine of N=5 celiacs activated an IFN-γ response when challenged with deamidated beer polypeptides.
authors.affiliation: Istituto di Scienze dell'Alimentazione (ISA) - CNR, Via Roma 64, 83100 Avellino, Italy. picariello@isa.cnr.it
authors.forename: Gianluca
authors.initials: G
authors.lastname: Picariello
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Mamone
authors.forename: Chiara
authors.initials: C
authors.lastname: Nitride
authors.forename: Francesco
authors.initials: F
authors.lastname: Addeo
authors.forename: Alessandra
authors.initials: A
authors.lastname: Camarca
authors.forename: Immacolata
authors.initials: I
authors.lastname: Vocca
authors.forename: Carmen
authors.initials: C
authors.lastname: Gianfrani
authors.forename: Pasquale
authors.initials: P
authors.lastname: Ferranti
mesh_terms: D000293:Adolescent; D000328:Adult; D000485:Allergens / Q000032:analysis*; D000595:Amino Acid Sequence; D001515:Beer / Q000032:analysis*; D002446:Celiac Disease / Q000276:immunology; D002648:Child; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid; D005983:Glutens / Q000032:analysis / Q000276:immunology*; D001467:Hordeum / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000276:immunology*; D010940:Plant Proteins / Q000032:analysis; D055315:Prolamins / Q000032:analysis; D020543:Proteome / Q000032:analysis*; D029701:Saccharomyces cerevisiae Proteins / Q000276:immunology; D021241:Spectrometry, Mass, Electrospray Ionization; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D000485:Allergens; D010455:Peptides; D010940:Plant Proteins; D055315:Prolamins; D020543:Proteome; D029701:Saccharomyces cerevisiae Proteins; D005983:Glutens