title: In vitro gastrointestinal digestion of the major peach allergen Pru p 3, a lipid transfer protein: molecular characterization of the products and assessment of their IgE binding abilities.
abstract: A simulated gastrointestinal digestion has been carried out on purified peach lipid transfer protein, one of the main allergens among the population of the Mediterranean area and the major allergen of peach allergic patients. The percentage of intact protein, after extensive digestion, measured by comparison with a non-digestible peptide analogue used as internal standard, was found to be about one-third of the original protein content. The peptides formed in digested fraction were characterized by means of LC/MS. The products of the digestion essentially derived from trypsin action, whereas the protein appeared to be resistant to pepsin and chymotrypsin. The identified peptides could be classified as low molecular weight and high molecular weight peptides. The latter consisted of the full protein, with the disulfide bridges still intact, deprived of the smaller peptides. The different digestion products, including the high and low molecular weight peptides, were purified by LC and assessed, together with the intact protein, by dot-blot analysis with sera of allergic patients, allowing to estimate their potential allergenicity. The intact protein and the high molecular weight peptides were found to be recognized by patients' sera, whereas the small peptides were found to be not reactive.
authors.affiliation: Department of Organic and Industrial Chemistry, University of Parma, Parma, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Cavatorta
authors.forename: Stefano
authors.initials: S
authors.lastname: Sforza
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Aquino
authors.forename: Gianni
authors.initials: G
authors.lastname: Galaverna
authors.forename: Arnaldo
authors.initials: A
authors.lastname: Dossena
authors.forename: Elide Anna
authors.initials: EA
authors.lastname: Pastorello
authors.forename: Rosangela
authors.initials: R
authors.lastname: Marchelli
mesh_terms: D000485:Allergens / Q000276:immunology* / Q000378:metabolism*; D000595:Amino Acid Sequence; D052179:Antigens, Plant / Q000276:immunology / Q000378:metabolism; D002352:Carrier Proteins / Q000276:immunology / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004044:Dietary Proteins / Q000276:immunology / Q000378:metabolism; D004063:Digestion; D005512:Food Hypersensitivity / Q000097:blood / Q000276:immunology; D005638:Fruit / Q000737:chemistry / Q000276:immunology; D006801:Humans; D006868:Hydrolysis; D007073:Immunoglobulin E / Q000276:immunology*; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry* / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D010940:Plant Proteins / Q000276:immunology / Q000378:metabolism; D011492:Protein Hydrolysates / Q000737:chemistry; D027861:Prunus / Q000737:chemistry / Q000276:immunology*; D021241:Spectrometry, Mass, Electrospray Ionization; D013379:Substrate Specificity; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D002352:Carrier Proteins; D004044:Dietary Proteins; D010446:Peptide Fragments; D010940:Plant Proteins; D011492:Protein Hydrolysates; C463181:Pru p 3 allergen; C511920:lipid transfer proteins, plant; D007073:Immunoglobulin E; D014357:Trypsin