Molecular characterization of a 10-kDa buckwheat molecule reactive to allergic patients' IgE.
BACKGROUND
Using the sera from buckwheat (BW)-allergic patients, several putative causative molecules were reported. However, few molecules were determined on the molecular structure. We demonstrated in 2000 that the major allergen with 24 kDa (BW24KD) is a legumin-like storage protein.


OBJECTIVE
The aim of this study was to isolate and characterize further a major allergen with 10 kDa by molecular cloning.


RESULTS
Buckwheat allergens were identified by immunoblotting analysis using sera from 14 allergic and two nonallergic individuals. We identified a protein with 10 kDa (BW10KD) that reacted with immunoglobulin E (IgE) more strongly than with IgG and IgA in 57% of the allergic patients but not with IgE in nonallergic individuals. Analyses were performed by N-terminal amino acid sequencing and molecular cloning. Physiological significance was assessed by an immunoblotting experiment showing that the reactivity of an allergic patient's serum IgE to BW10KD was competitively inhibited by natural BW extracts.


CONCLUSIONS
Molecular cloning experiments indicated that BW10KD as a BW allergen was a member of the 2S-albumin multigene family.
Matsumoto
R
R
Department of Bioengineering, Faculty of Engineering, Kagoshima University, Kagoshima, Japan.
Fujino
K
K
Nagata
Y
Y
Hashiguchi
S
S
Ito
Y
Y
Aihara
Y
Y
Takahashi
Y
Y
Maeda
K
K
Sugimura
K
K
D000293:Adolescent; D000328:Adult; D000368:Aged; D000485:Allergens / Q000737:chemistry / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D002648:Child; D002675:Child, Preschool; D019612:Fagopyrum / Q000737:chemistry / Q000276:immunology*; D005260:Female; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology*; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010936:Plant Extracts / Q000737:chemistry
D000485:Allergens; D010936:Plant Extracts; D007073:Immunoglobulin E