6.5.1 Molecular Biological Properties
 

Soybean Vacuolar Protein	References
Allergen Nomenclature Gly m Bd 30 K	(1) Ogawa et al. 1991
Molecular Mass  monomer: 34 kDa (1)*, 32 kDa (2)*, 30 kDa (SDS-PAGE) (3) oligomer: 3000 kDa (Gel filtration) (2) precursor: 46-47 kDa (1, 4) *34 kDa polypeptide is processed during seedling growth to a 32 kDa polypeptide by the removal of an N-terminal decapeptide (1)	(1) Kalinski et al. 1990, 1992 (2) Ogawa et al. 1993 (3) Helm et al. 1998 (4) Ji et al. 1998
Isoelectric Point pI 4.5 (oligomer)	(1) Ogawa et al. 1993
Amino Acid Sequence, mRNA, and cDNA Proteins Gly m Bd 30K* P34* SWISS-PROT: P22895 TrEMBL:  O64458   GenBank: AB013289 J05560 Amino Acids 379 (2) 379 (1) mRNA precursor   1287 bp (1) cDNA precursor 6675 bp (2)    * Gly m Bd 30K and P34 are identical (Ogawa et al. 1995)	(1) Kalinski et al. 1990, 1992 (2) Takano et al. 1995
recombinant Protein Expression in Escherichia coli: Isolation of protein coding region from clone P34 by PCR technique, subcloned into pet-24(b)+ vector and expression in E.coli (2) Expression in insect cells: 46-, 34-, and 32-kDa forms (1)	(1) Ji et al. 1998 (2) Helm et al. 2000
Posttranslational Modifications Glycosylation: Carbohydrate composition: Man, GlcNAc, Fuc, and Xyl (3:2:1:1, molar ratio) (1) N-Glycosylation site: Asn-170 (mature protein) (1)	(1) Bando et al. 1996
Biological Function cysteine protease (papain superfamily)	(1) Kalinski et al. 1992
Sequence Homology Der p 1, a house dust mite allergen: 30% identity (1), 54% similarity (2)	(1) Ogawa et al. 1993 (2) Helm et al. 1998
Localization Seed protein storage vacuoles in soybean cotyledons (electron microscopic immunolocalization, mAb (1, 2), patients' IgE (2))	(1) Kalinski et al. 1992 (2) Helm et al. 1998
Other Properties Binding to alpha'- and alpha-subunits of beta-conglycinin (1) Binding of syringolides (which trigger defense responses) (2)	(1) Samoto et al. 1996 (2) Ji et al. 1998

 6.5.2 Allergenic Properties
 

Soybean Vacuolar Protein	References
Frequency of Sensitization IgE-binding to Gly m Bd 30K in 65% of patients (1)	(1) see 6.1 Sensitization to Soybean Allergens
B-Cell Epitopes IgE binding sites located on Gly m Bd 30K (the precursor sequence): Peptides Positivity in Patients aa 3-12 (synthetic peptide) + (1), 67% (2) aa 100-110 (synthetic peptide) + (1), 50% (2) aa 229-238 (synthetic peptide) + (1), 58% (2) aa 299-308 (synthetic peptide) + (1), 67% (2) aa 331-340 (synthetic peptide) + (1), 75% (2) dot / immunoblot (SPOTs membrane technique) (1) 4 patients with soybean allergy (DBPCFC) (2) additional 8 patients with soybean allergy (DBPCFC)	(1) Helm et al. 1998 (2) Helm et al. 2000
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 100-110 (synthetic peptide) a) S105A b) K106A a) 0% (1) b) 0% (1) aa 331-340 (synthetic peptide) a) I315A b) Q316A c) R317A a) 66% (1) b) 66% (1) c) 33% (1) dot / immunoblot (SPOTs membrane technique) (1) 6 patients with soybean allergy (DBPCFC) and positive IgE-binding to non-mutated peptide	(1) Helm et al. 2000
Alteration of Allergenicity galactomannan treatment (Maillard reaction): complete reduction of allergenicity (a) (1) (a) dot / immunoblot	(1) Babiker et al. 1998