Open AccessIdentification of major wheat allergens by means of the Escherichia coli expression system
Author(s) -
Maruyama Nobuyuki,
Ichise Keiko,
Katsube Tomoyuki,
Kishimoto Toru,
Kawase Shinichiro,
Matsumura Yasuki,
Takeuchi Yoshihiro,
Sawada Tadashi,
Utsumi Shigeru
Publication year - 1998
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1327.1998.2550739.x
Subject(s) - glutenin , gliadin , prolamin , chemistry , recombinant dna , wheat allergy , escherichia coli , epitope , glutelin , molecular mass , biochemistry , microbiology and biotechnology , storage protein , gluten , gene , biology , antigen , immunoglobulin e , antibody , genetics , protein subunit , enzyme
Wheat proteins were fractionated into salt‐soluble, glutenin‐rich, and gliadin‐rich fractions. Reactivities of these protein fractions with sera of patients with wheat‐associated allergies were examined under various conditions. The relative reactivity of the fractions was generally in the order glutenin‐rich > gliadin‐rich @WR salt‐soluble fractions, although their reactivities were variable among patients and among the reaction conditions, indicating that the kind, the number and the epitope of allergens were variable among patients. To identify major allergens, α‐, γ‐ and ω‐gliadin, and low‐molecular‐mass (LMM)‐ and high‐molecular‐mass (HMM)‐glutenin genes were expressed in Escherichia coli by means of a pET vector. Recombinant gliadins and glutenins were partially purfied on the basis of the solubilities of prolamin and glutelin. The partially purified recombinant proteins were reacted with the patients' sera. LMM glutenin containing many Gln‐Gln‐Gln‐Pro‐Pro motifs, which was identified to be IgE‐binding epitope [Tanabe, S., Arai, S., Yanagihara, Y., Mita, H., Takahashi, K. & Watanabe, M. (1996) Biochem. Biophys. Res. Commun. 219 , 290−293], exhibited the highest reactivity. The next highest reactivities were observed on α‐gliadin and γ‐gliadin, which had not been identified as allergens.