PMID- 15714473
OWN - NLM
STAT- MEDLINE
DCOM- 20050719
LR  - 20191210
IS  - 1615-9853 (Print)
IS  - 1615-9853 (Linking)
VI  - 5
IP  - 3
DP  - 2005 Feb
TI  - Analysis of the composition of an immunoglobulin E reactive high molecular weight 
      protein complex of peanut extract containing Ara h 1 and Ara h 3/4.
PG  - 675-86
AB  - Peanuts (Arachis hypogaea) contain some of the most potent food allergens. In 
      recent years an increasing prevalence of peanut allergies both in children and 
      adults has been observed in the USA and in Europe. In vitro identification and 
      characterization of allergens including those from peanut have been frequently 
      performed by Western blotting. However this method may alter the immunoglobulin E 
      (IgE) antibody reactivity since the proteins are denatured by detergent treatment 
      and/or reduction of disulfide bonds by reducing reagents and does not answer the 
      question how peanut allergens interact with the human digestive apparatus and 
      immune system. Size exclusion chromatography of peanut extract shows that 
      approximately 90% of the total protein content is eluted as one peak in the 
      exclusion volume with a molecular mass of over 200 kDa. The proteins of this 
      fraction were analyzed by blue-native polyacrylamide gel electrophoresis (PAGE), 
      immunoblotting, two-dimensional PAGE and Western blotting. A complex of Ara h 1 
      (Acc. no. P43237), Ara h 3/4 (AAM46958), Ara h 3 (AAC63045), Ara h 4 (AF086821), 
      Gly 1 (AAG01363) and iso-Ara h 3 (AAT39430) was identified using patients' IgE 
      and allergen-specific monoclonal antibodies; N-terminal sequencing and 
      matrix-assisted laser desorption/ionisation-time of flight analysis verified 
      these findings. A comparison of the peanut allergen sequences of Ara h 3/4, Ara h 
      3, Ara h 4 and peanut trypsin inhibitor (AF487543) and the proteins Gly 1 and 
      iso-Ara h 3, not yet described as allergens, leads to the conclusion that these 
      proteins are isoallergens of each other. It was shown that these isoallergens are 
      post-translationally cleaved and held together by disulfide bonds in accordance 
      to the 11S plant seed storage proteins signature.
FAU - Boldt, Andrea
AU  - Boldt A
AD  - Research Center Borstel, Borstel, Germany.
FAU - Fortunato, Donatella
AU  - Fortunato D
FAU - Conti, Amedeo
AU  - Conti A
FAU - Petersen, Arnd
AU  - Petersen A
FAU - Ballmer-Weber, Barbara
AU  - Ballmer-Weber B
FAU - Lepp, Ute
AU  - Lepp U
FAU - Reese, Gerald
AU  - Reese G
FAU - Becker, Wolf-Meinhard
AU  - Becker WM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - Proteomics
JT  - Proteomics
JID - 101092707
RN  - 0 (Allergens)
RN  - 0 (Antigens, Plant)
RN  - 0 (Ara h 1 protein, Arachis hypogaea)
RN  - 0 (Ara h 4 allergen, Arachis hypogaea)
RN  - 0 (Glycoproteins)
RN  - 0 (Membrane Proteins)
RN  - 0 (Plant Extracts)
RN  - 0 (Plant Proteins)
RN  - 0 (Seed Storage Proteins)
RN  - 0 (allergen Ara h3)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Allergens/*analysis/immunology
MH  - Amino Acid Sequence
MH  - Antigens, Plant
MH  - *Arachis
MH  - Chromatography, Gel
MH  - Electrophoresis, Gel, Two-Dimensional
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Glycoproteins/*analysis/immunology
MH  - Humans
MH  - Immunoblotting
MH  - Immunoglobulin E/*immunology
MH  - Membrane Proteins
MH  - Molecular Sequence Data
MH  - Molecular Weight
MH  - Peptide Mapping
MH  - Plant Extracts/chemistry/immunology
MH  - Plant Proteins/*analysis/immunology
MH  - Seed Storage Proteins
MH  - Sequence Analysis, Protein
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
EDAT- 2005/02/17 09:00
MHDA- 2005/07/20 09:00
CRDT- 2005/02/17 09:00
PHST- 2005/02/17 09:00 [pubmed]
PHST- 2005/07/20 09:00 [medline]
PHST- 2005/02/17 09:00 [entrez]
AID - 10.1002/pmic.200401150 [doi]
PST - ppublish
SO  - Proteomics. 2005 Feb;5(3):675-86. doi: 10.1002/pmic.200401150.