Mutational analysis of the IgE-binding epitopes of P34/Gly m Bd 30K.
BACKGROUND
Peanuts and soybeans are 2 foods that have been shown to be responsible for many atopic disorders. Because of their nutritional benefit, soybean proteins are now being used increasingly in a number of food products. Previous studies have documented multiple allergens in soybean extracts, including glycinin, beta-conglycinin, and the P34/Gly m Bd 30K protein.


OBJECTIVE
Our overall goal was to identify soybean-specific allergens to begin to understand molecular and immunochemical characteristics of legume proteins. The specific aim of the current investigation was to identify the essential amino acid residues necessary for IgE binding in the 5 distinct immunodominant epitopes of P34/Gly m Bd 30K.


METHODS
Serum IgE from 6 clinically sensitive soybean-allergic individuals was used to identify P34/Gly m Bd 30K in the native and single amino acid substituted peptides with use of the SPOTS peptide synthesis technique to determine critical amino acids required for IgE binding.


RESULTS
The intensity of IgE binding and epitope recognition by serum IgE from the individuals varied substantially. With use of serum from 6 clinically soybean-sensitive individuals, 2 of the 5 immunodominant epitopes could be mutagenized to non-IgE binding peptides.


CONCLUSIONS
Single-site amino acid substitution of the 5 immunodominant epitopes of Gly m Bd 30K with alanine revealed that IgE binding could be reduced or eliminated in epitopes 6 and 16 in the serum obtained from 6 soybean-sensitive patients.
Helm
R M
RM
Department of Pediatrics, University of Arkansas for Medical Sciences, Arkansas Children's Nutrition Center, Little Rock, AR 72202, USA.
Cockrell
G
G
Connaughton
C
C
West
C M
CM
Herman
E
E
Sampson
H A
HA
Bannon
G A
GA
Burks
A W
AW
D000485:Allergens / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001666:Binding Sites, Antibody; D004252:DNA Mutational Analysis; D018744:DNA, Plant / Q000276:immunology / Q000378:metabolism*; D004311:Double-Blind Method; D006801:Humans; D016056:Immunodominant Epitopes / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D007073:Immunoglobulin E / Q000097:blood / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010940:Plant Proteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D030262:Soybean Proteins; D013025:Glycine max / Q000276:immunology
D000485:Allergens; D052179:Antigens, Plant; D018744:DNA, Plant; C103236:Gly m BD 30K protein, Glycine max; D016056:Immunodominant Epitopes; D010940:Plant Proteins; D011994:Recombinant Proteins; D030262:Soybean Proteins; D007073:Immunoglobulin E