PMID- 9186485
OWN - NLM
STAT- MEDLINE
DCOM- 19970710
LR  - 20151119
IS  - 0003-9861 (Print)
IS  - 0003-9861 (Linking)
VI  - 342
IP  - 2
DP  - 1997 Jun 15
TI  - Identification and mutational analysis of the immunodominant IgE binding epitopes 
      of the major peanut allergen Ara h 2.
PG  - 244-53
AB  - A major peanut allergen, Ara h 2, is recognized by serum IgE from > 90% of 
      patients with peanut hypersensitivity. Biochemical characterization of this 
      allergen indicates that it is a glycoprotein of approximately 17.5 kDa. Using 
      N-terminal amino acid sequence data from purified Ara h 2, oligonucleotide 
      primers were synthesized and used to identify a clone (741 bp) from a peanut cDNA 
      library. This clone was capable of encoding a 17.5-kDa protein with homology to 
      the conglutin family of seed storage proteins. The major linear immunoglobulin E 
      (IgE)-binding epitopes of this allergen were mapped using overlapping peptides 
      synthesized on an activated cellulose membrane and pooled serum IgE from 15 
      peanut-sensitive patients. Ten IgE-binding epitopes were identified, distributed 
      throughout the length of the Ara h 2 protein. Sixty-three percent of the amino 
      acids represented in the epitopes were either polar uncharged or apolar residues. 
      In an effort to determine which, if any, of the 10 epitopes were recognized by 
      the majority of patients with peanut hypersensitivity, each set of 10 peptides 
      was probed individually with serum IgE from 10 different patients. All of the 
      patient sera tested recognized multiple epitopes. Three epitopes (aa27-36, 
      aa57-66, and aa65-74) were recognized by all patients tested. In addition, these 
      three peptides bound more IgE than all the other epitopes combined, indicating 
      that they are the immunodominant epitopes of the Ara h 2 protein. Mutational 
      analysis of the Ara h 2 epitopes indicate that single amino acid changes result 
      in loss of IgE binding. Two epitopes in region aa57-74 contained the amino acid 
      sequence DPYSP that appears to be necessary for IgE binding. These results may 
      allow for the design of improved diagnostic and therapeutic approaches to peanut 
      hypersensitivity.
FAU - Stanley, J S
AU  - Stanley JS
AD  - Department of Biochemistry & Molecular Biology, University of Arkansas for 
      Medical Sciences, Arkansas Children's Hospital Research Institute, Little Rock 
      72205, USA.
FAU - King, N
AU  - King N
FAU - Burks, A W
AU  - Burks AW
FAU - Huang, S K
AU  - Huang SK
FAU - Sampson, H
AU  - Sampson H
FAU - Cockrell, G
AU  - Cockrell G
FAU - Helm, R M
AU  - Helm RM
FAU - West, C M
AU  - West CM
FAU - Bannon, G A
AU  - Bannon GA
LA  - eng
GR  - R01-AI33596/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Arch Biochem Biophys
JT  - Archives of biochemistry and biophysics
JID - 0372430
RN  - 0 (2S Albumins, Plant)
RN  - 0 (Allergens)
RN  - 0 (Antigens, Plant)
RN  - 0 (Ara h II protein, Arachis hypogaea)
RN  - 0 (Epitopes)
RN  - 0 (Glycoproteins)
RN  - 0 (Immunodominant Epitopes)
RN  - 0 (Peptide Fragments)
RN  - 0 (Plant Proteins)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - 2S Albumins, Plant
MH  - Adult
MH  - Allergens/chemistry/*immunology
MH  - Amino Acid Sequence
MH  - Antigens, Plant
MH  - Arachis/*immunology
MH  - Base Sequence
MH  - Binding Sites, Antibody
MH  - Cloning, Molecular
MH  - DNA Mutational Analysis
MH  - Epitope Mapping
MH  - Epitopes/chemistry/immunology
MH  - Food Hypersensitivity/immunology
MH  - Glycoproteins/chemistry/*immunology
MH  - Humans
MH  - Immunodominant Epitopes
MH  - Immunoglobulin E/*immunology
MH  - Molecular Sequence Data
MH  - Peptide Fragments/chemical synthesis/chemistry/immunology
MH  - Plant Proteins
MH  - Sequence Homology, Amino Acid
EDAT- 1997/06/15 00:00
MHDA- 1997/06/15 00:01
CRDT- 1997/06/15 00:00
PHST- 1997/06/15 00:00 [pubmed]
PHST- 1997/06/15 00:01 [medline]
PHST- 1997/06/15 00:00 [entrez]
AID - S0003-9861(97)99998-7 [pii]
AID - 10.1006/abbi.1997.9998 [doi]
PST - ppublish
SO  - Arch Biochem Biophys. 1997 Jun 15;342(2):244-53. doi: 10.1006/abbi.1997.9998.