Pyr c 1, the major allergen from pear (Pyrus communis), is a new member of the Bet v 1 allergen family

Karamloo, Fariba; Scheurer, Stephan; Wangorsch, Andrea; May, Sibylle; Haustein, Dieter; Vieths, Stefan

doi:10.1016/s0378-4347(01)00087-1

Cited by 67 publications

(37 citation statements)

References 36 publications

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“…The amino-acid sequence of the P-loop of Bet v 1 is highly conserved in Bet v 1-related food allergens, such as Pru av 1, Pyr c 1 [44] from pear (Pyrus communis), Mal d 1 [45] from apple (Malus domestica) and Api g 1.0201 [13]. In contrast, Api g 1.0101 [12] lacks Leu 44 and the negatively charged Glu 45 is substituted by the positively charged Lys 44 (Figure 1).…”

Section: Expression and Purification Of Recombinant Pru Av 1 Api G 1mentioning

confidence: 99%

“…In contrast, Api g 1.0101 [12] lacks Leu 44 and the negatively charged Glu 45 is substituted by the positively charged Lys 44 (Figure 1). Furthermore, the C-terminus of Api g 1.0101 is truncated by five amino acids.…”

Section: Expression and Purification Of Recombinant Pru Av 1 Api G 1mentioning

confidence: 99%

“…Furthermore, the C-terminus of Api g 1.0101 is truncated by five amino acids. Hence, the amino acids Glu 45 , Ser 112 and 155-159 of Pru av 1 and Lys 44 of Api g 1.0101 were selected for site-directed mutagenesis and recombinant proteins were expressed in Escherichia coli. SDS/PAGE analysis revealed that all recombinant cherry proteins were obtained with a high degree of purity (> 98 %; Figure 2), except for Pru av 1 Ala 112 with a purity of approx.…”

Section: Expression and Purification Of Recombinant Pru Av 1 Api G 1mentioning

confidence: 99%

“…Pru av 1 Arg 45 underwent proteolysis after expression and was therefore not investigated any further. Api g 1.0101 wt and Glu 44 were also prepared with high purity, analysed by SDS/PAGE and Coomassie Blue staining (results not shown). …”

Section: Expression and Purification Of Recombinant Pru Av 1 Api G 1mentioning

confidence: 99%

“…68 % for a subset of cherry allergic patients. The experiments were extended by comparing the IgE reactivity of the celery allergens Api g 1.0101 wt and Glu 44 . Nine out of 15 celery allergic patients were sensitized to Api g 1.0101.…”

Section: Analysis Of the Ige Reactivitymentioning

confidence: 99%

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Mutational epitope analysis of Pru av 1 and Api g 1, the major allergens of cherry (Prunus avium) and celery (Apium graveolens): correlating IgE reactivity with three-dimensional structure

Neudecker

Lehmann

Nerkamp

et al. 2003

Biochemical Journal

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112

106

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Birch pollinosis is often accompanied by adverse reactions to food due to pollen-allergen specific IgE cross-reacting with homologous food allergens. The tertiary structure of Pru av 1, the major cherry (Prunus avium) allergen, for example, is nearly identical with Bet v 1, the major birch (Betula verrucosa) pollen allergen. In order to define cross-reactive IgE epitopes, we generated and analysed mutants of Pru av 1 and Api g 1.0101, the major celery (Apium graveolens) allergen, by immunoblotting, EAST (enzyme allergosorbent test), CD and NMR spectroscopy. The mutation of Glu 45 to Trp 45 in the P-loop region, a known IgE epitope of Bet v 1, significantly reduced IgE binding to Pru av 1 in a subgroup of cherry-allergic patients. The backbone conformation of Pru av 1 wild-type is conserved in the three-dimensional structure of Pru av 1 Trp 45 , demonstrating that the side chain of Glu 45 is involved in a cross-reactive IgE epitope. Accordingly, for a subgroup of celery-allergic patients, IgE binding to the homologous celery allergen Api g 1.0101 was enhanced by the mutation of Lys 44 to Glu. The almost complete loss of IgE reactivity to the Pru av 1 Pro 112 mutant is due to disruption of its tertiary structure. Neither the mutation Ala 112 nor deletion of the Cterminal residues 155-159 influenced IgE binding to Pru av 1. In conclusion, the structure of the P-loop partially explains the crossreactivity pattern, and modulation of IgE-binding by site-directed mutagenesis is a promising approach to develop hypo-allergenic variants for patient-tailored specific immunotherapy.

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