7.4.1 Molecular Biological Properties
 

Glycinin	References
Allergen Nomenclature  Ara h 3 / Ara h 4	(1) Larsen & Lowenstein 1999
Isoallergens and Variants At least 2 isoallergens designated Ara h 3 and Ara h 4 sharing 91.3% aa sequence identity (not known at the time of approval of names by the IUIS Allergen Nomenclature Sub- Committee) (1)	(1) Kleber-Janke et al. 1999
Molecular Mass SDS-PAGE: 14 kDa (1)*, 57 kDa (rAra h 3) (2) calculated 35.9 kDa (3), 61.0 kDa (full length clone, 3) *N-terminal breakdown product (?)	(1) Burks et al. 1998b (2) Rabjohn et al. 1999 (3) Kleber-Janke et al. 1999
Isoelectric Point  Calculated pI 5.5 (1)	(1) Kleber-Janke et al. 1999
Amino Acid Sequence, mRNA, and cDNA   Ara h 3 (1) Ara h 4 (2) SWISS-PROT:     GenBank: AF093541 AF086821 PIR:     Amino acids 510 residues (1) 315 residues (2)* mRNA 1524 bp 1853 bp cDNA 1530 bp (1) 1170 bp (2)* Gene     *full lenght cDNA clone 1872 bp, 530 aa, 61.0 kDa (2)	(1) Rabjohn et al. 1999 (2) Kleber-Janke et al. 1999
Recombinant Protein cDNA expression library: Isolation of Ara h 3 mRNA by PCR  (1) Phage surface display of functional cDNA expression products, selection of cDNA clones on microtiter wells coated with serum IgE from peanut sensitive patients (pJuFo cloning system) (2) Expression in Escherichia coli: Subcloning into pDS56/RBSII vector and expression of recombinant Ara h 4 in E.coli M15 cells (2) Expression in bacterial cells: Expression of recombinant Ara h 3 by using procaryotic expression vector pET24 plasmid (1)	(1) Rabjohn et al. 1999 (2) Kleber-Janke et al. 1999
Biological Function 11S seed storage protein (1)	(1) Rabjohn et al. 1999
Sequence Homology Glycinins (11S-storage proteins) from soybean and pea: aa 62-72% identity with Ara h 3 (1) Glycinin (11S-storage proteins) from soybean: aa 56% identity with Ara h 4 (2)	(1) Rabjohn et al. 1999 (2) Kleber-Janke et al. 1999

7.4.2 Allergenic Properties
 

Glycinin	References
Frequency of Sensitization IgE-binding to Ara h 3 in 44-53% of patients (1)	(1) see 7.1 Sensitization to Peanut Allergens
IgE binding to rAra h 3 44% of 18 peanut allergic patients showed IgE- binding to recombinant Ara h 3 (SDS-PAGE immunoblot) (1)	(1) Rabjohn et al. 1999
B-Cell Epitopes IgE binding sites located on Ara h 3: Peptides Positivity in Patients aa 33-47 (synthetic peptide) 25% (1) aa 240-254 (synthetic peptide) 38% (1) aa 279-293 (synthetic peptide) 100% (1) aa 303-317 (synthetic peptide) 38% (1) dot / immunoblot (SPOTs membrane technique) (1) Sera with Ara h 3 specific IgE from 8 patients with peanut allergy	(1) Rabjohn et al. 1999
Mutational Analysis of B-Cell Epitopes Critical aa residues for IgE binding identified by single site aa substitution: Peptides aa Substitution Positivity in Patients aa 33-47 (synthetic peptide) a) P38A b) N39A - (1) aa 240-254 (synthetic peptide) a) F244A b) F247A c) F250A d) L251A - (1) aa 279-293 (synthetic peptide) a) L285A b) I287A c) L288A d) P290A - (1) aa 303-317 (synthetic peptide) a) E308A b) Y309A c) D310A d) E311A - (1) dot / immunoblot (SPOTs membrane technique) (1) Serum pooled from 8 patients with peanut allergy	(1) Rabjohn et al. 1999
Types of Critical Amino Acids 68% of amino acids from 4 epitopes were either polar uncharged or apolar residues (1)	(1) Rabjohn et al. 1999