title: Absolute quantification of Pru av 2 in sweet cherry fruit by liquid chromatography/tandem mass spectrometry with the use of a stable isotope-labelled peptide.
abstract: Pru av 2, a pathogenesis-related (PR) protein present in the sweet cherry (Prunus avium L.) fruit, is the principal allergen of cherry and one of the chief causes of pollen food syndrome (oral allergy syndrome). In this study, a quantitative assay for this protein was developed with the use of the protein absolute quantification (AQUA) method, which consists of liquid chromatography/tandem mass spectrometry (LC/MS/MS) employing TGC[CAM]STDASGK[(13)C6,(15)N2], a stable isotope-labelled internal standard (SIIS) peptide. This assay gave a linear relationship (r(2)>0.99) in a concentration range (2.3-600fmol/μL), and the overall coefficient of variation (CV) for multiple tests was 14.6%. Thus, the contents of this allergenic protein in sweet cherry products could be determined using this assay. This assay should be valuable for allergological investigations of Pru av 2 in sweet cherry and detection of protein contamination in foods.
authors.affiliation: National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. Electronic address: ippoushi@affrc.go.jp.
authors.forename: Katsunari
authors.initials: K
authors.lastname: Ippoushi
authors.affiliation: National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
authors.forename: Motoe
authors.initials: M
authors.lastname: Sasanuma
authors.affiliation: National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Oike
authors.affiliation: National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
authors.forename: Masuko
authors.initials: M
authors.lastname: Kobori
authors.affiliation: National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
authors.forename: Mari
authors.initials: M
authors.lastname: Maeda-Yamamoto
mesh_terms: D000485:Allergens / Q000737:chemistry* / Q000276:immunology; D000595:Amino Acid Sequence; D052179:Antigens, Plant / Q000737:chemistry* / Q000276:immunology; D002247:Carbon Isotopes / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005512:Food Hypersensitivity / Q000276:immunology; D005638:Fruit / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D007553:Isotope Labeling; D009587:Nitrogen Isotopes / Q000032:analysis; D010455:Peptides / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry / Q000276:immunology; D000068220:Prunus avium / Q000737:chemistry* / Q000276:immunology; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D010455:Peptides; D010940:Plant Proteins; C510168:Pru av 2 allergen, Prunus avium
keywords: Absolute quantification; Liquid chromatography/tandem mass spectrometry; Pru av 2; Prunus avium L.; Stable isotope-labelled peptide; Sweet cherry; Trypsin