Cloning of the minor allergen Api g 4 profilin from celery (<i>Apium graveolens</i>) and its cross‐reactivity with birch pollen profilin Bet v 2

Scheurer, Stephan; Wangorsch, Andrea; Haustein, Dieter; Vieths, Stefan

doi:10.1046/j.1365-2222.2000.00821.x

Cited by 91 publications

(74 citation statements)

References 38 publications

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“…Recombinant Pru av 1 wt, Trp 45 , Ala 112 , Pro 112 and 155-159 were coupled to CNBractivated filter paper discs (Hycor, Kassel, Germany) at a protein concentration of 1.5 µg/ml [21]. Dose-related EAST inhibition experiments with 0.5 µg of recombinant Pru av 1 immobilized on paper discs were performed as described previously [22]. Pooled serum samples were diluted 1:2 and subsequently pre-incubated with dilutions of recombinant Pru av 1 wt, Trp 45 …”

Section: East and East Inhibitionmentioning

confidence: 99%

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

110

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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Section: East and East Inhibitionmentioning

confidence: 99%

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

110

106

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“…Detection of binding of birch pollen profilin-specific IgE to reactive epitopes in PVDF membrane-bound recombinant birch profilin was performed as previously described (5,(17)(18)(19). Briefly, native or ␣-chymasecleaved purified recombinant birch profilin (BIOMAY) was electrophoresed onto 12% bis-Tris polyacrylamide gels and transferred to PVDF as described above.…”

Section: Ige Immunoblottingmentioning

confidence: 99%

“…The use of control serum from nonallergic individuals establishes the specificity of the binding of IgE present in allergic serum, while preadsorption of allergic sera with purified allergens in IgE inhibition experiments may identify cross-sensitization of individuals to allergens in unrelated trees, weeds, or grasses (5,(17)(18)(19). To determine the effect of ␣-chymase hydrolysis of birch profilin on binding of allergenspecific IgE, immunoblotting of native or ␣-chymase-cleaved birch profilin was performed using serum from individuals allergic to birch pollen.…”

Section: Ige Immunoblotmentioning

confidence: 99%

Mast Cell α-Chymase Reduces IgE Recognition of Birch Pollen Profilin by Cleaving Antibody-Binding Epitopes

Mellon

Frank

Fang

2002

The Journal of Immunology

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In sensitized individuals birch pollen induces an allergic response characterized by IgE-dependent mast cell degranulation of mediators, such as α-chymase and other serine proteases. In birch and other plant pollens, a major allergen is profilin. In mammals, profilin homologues are found in an intracellular form bound to cytoskeletal or cytosolic proteins or in a secreted form that may initiate signal transduction. IgE specific to birch profilin also binds human profilin I. This cross-reactivity between airborne and endogenous proteins may help to sustain allergy symptoms. The current work demonstrates that cultured mast cells constitutively secrete profilin I, which is susceptible to degranulation-dependent proteolysis. Coincubation of chymase-rich BR mastocytoma cells with Ala-Ala-Pro-Phe-chloromethylketone (a chymase inhibitor) blocks profilin cleavage, which does not occur in degranulated HMC-1 mast cells, which are rich in tryptase, but chymase deficient. These data implicate chymase as the serine protease cleaving secreted mast cell profilin. Sequencing of chymase-cleaved profilins reveals hydrolysis at Tyr6-Val7 and Trp35-Ala36 in birch profilin and at Trp32-Ala33 in human profilin, with all sites lying within IgE-reactive epitopes. IgE immunoblotting studies with sera from birch pollen-allergic individuals demonstrate that cleavage by chymase attenuates binding of birch profilin to IgE. Thus, destruction of IgE-binding epitopes by exocytosed chymase may limit further mast cell activation by this class of common plant allergens, thereby limiting the allergic responses in sensitized individuals.

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“…The basic biological functions of profilins have been attributed to cell elongation, cell shape maintenance, and flowering (Ramachandran et al 2000), seedling development (McKinney et al 2001) and pollen tube growth (McKenna et al 2004). Apart from pollen, many fruits contain profilins and their allergenic potency has been frequently reported (Scheurer et al 2000(Scheurer et al , 2001Wensing et al 2002;Asero et al 2003;Rodriguez-Perez et al 2003;Westphal et al 2004). When we started this research, three distinct mRNA sequences (AF129426, AF129427 and AF129428) encoding three isoallergens of apple profilin were available in the GenBank/EMBL nucleotide database.…”

mentioning

confidence: 99%

Genomic characterization and linkage mapping of the apple allergen genes Mal d 2 (thaumatin-like protein) and Mal d 4 (profilin)

et al. 2005

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Cloning of the minor allergen Api g 4 profilin from celery (Apium graveolens) and its cross‐reactivity with birch pollen profilin Bet v 2

Cited by 91 publications

References 38 publications

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

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

Mast Cell α-Chymase Reduces IgE Recognition of Birch Pollen Profilin by Cleaving Antibody-Binding Epitopes

Genomic characterization and linkage mapping of the apple allergen genes Mal d 2 (thaumatin-like protein) and Mal d 4 (profilin)

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