Biochemical and structural analysis of the IgE binding sites on ara h1, an abundant and highly allergenic peanut protein.
Allergy to peanut is a significant IgE-mediated health problem because of the high prevalence, potential severity, and chronicity of the reaction. Ara h1, an abundant peanut protein, is recognized by serum IgE from >90% of peanut-sensitive individuals. It has been shown to belong to the vicilin family of seed storage proteins and to contain 23 linear IgE binding epitopes. In this communication, we have determined the critical amino acids within each of the IgE binding epitopes of Ara h1 that are important for immunoglobulin binding. Surprisingly, substitution of a single amino acid within each of the epitopes led to loss of IgE binding. In addition, hydrophobic residues appeared to be most critical for IgE binding. The position of each of the IgE binding epitopes on a homology-based molecular model of Ara h1 showed that they were clustered into two main regions, despite their more even distribution in the primary sequence. Finally, we have shown that Ara h1 forms a stable trimer by the use of a reproducible fluorescence assay. This information will be important in studies designed to reduce the risk of peanut-induced anaphylaxis by lowering the IgE binding capacity of the allergen.
Shin
D S
DS
Department of Biochemistry & Molecular Biology, University of Arkansas for Medical Sciences, Arkansas Children's Hospital, Little Rock, Arkansas 72205, USA.
Compadre
C M
CM
Maleki
S J
SJ
Kopper
R A
RA
Sampson
H
H
Huang
S K
SK
Burks
A W
AW
Bannon
G A
GA
D000328:Adult; D000485:Allergens / Q000737:chemistry / Q000378:metabolism*; D000595:Amino Acid Sequence; D010367:Arachis / Q000276:immunology*; D001665:Binding Sites; D018360:Crystallography, X-Ray; D000939:Epitopes / Q000378:metabolism; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism*; D008969:Molecular Sequence Data; D010919:Placebos; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid
D000485:Allergens; D000939:Epitopes; D010919:Placebos; D007073:Immunoglobulin E