Allergy information for: Cherry (Prunus avium )

Clinical History

• Number of Studies:1-5
• Number of Patients:>50
• Symptoms:

  Several studies have reported symptoms of cherry allergy. They include oral allergy syndrome (OAS) with mild itching of the lips, tongue or throat in patients with reported allergy to cherry and a positive IgE CAP to birch pollen (Scheurer et al. 2001).

  Other symtoms less frequents are urticaria, angioedema, severe dyspnea, cough, and gastrointestinal symptoms accompanied by severe pain after cherry consumption. (Ballmer-Weber et al. 2002) (Scheurer et al. 2001).

Skin Prick Test

• Number of Studies:1-5
• Food/Type of allergen:

  Fresh fruit and commercial cherry extracts (Pastorello et al. 1994)

  Purified allergens (recombinant (r) Pru av 1; rPru av 4; and lipid transfer protein: rPru av 3), commercial cherry extacts and home-made extracts as follows. Fresh fruits were cut and homogenised in acetone/dry ice at -60 to -65°C and the mixture was stored overnight and cooled with dry ice. The resulting precipitates were washed twice with acetone (–60°C) and once with chilled acetone-diethyl ether (1:1, –60°C). The filter residue was lyophilized. 2 gr of the obtained acetone powder was extracted with 30 mL of PBS (0.01M potassium phosphate and 0.15 M NaCl (pH 7.4)). The suspension was centrifuged for 45 minutes at 20,000g, and the supernatant was filtered through a fiber glass prefilter (Ballmer-Weber et al. 2002).

• Protocol: (controls, definition of positive etc)

  Pastorello et al. 1994 used histamine dihydrochloride (10 mg/mL) as a positive control, and the glycerol-containing diluent of the prick solution as a negative control. A wheal graded at least 2+was regarded as positive.

  Ballmer-Weber et al. 2002 used histamine dihydrochloride (10 mg/mL) as a positive control, and the glycerol-containing diluent of the prick solution as a negative control. A wheal size of 7 mm² or greater was regarded as positive.

• Number of Patients:

  21 patients (Pastorello et al. 1994)

  79 subjects were included in the study: 24 Swiss patients (group 1) with a positive double-blind placebo-controlled food challenge result to cherries, 23 patients with birch pollen allergy but without cherry allergy (group 2), 23 nonatopic subjects (group 3), and 9 Spanish patients with a history of a cherry allergy (group 4) (Ballmer-Weber et al. 2002).

• Summary of Results:

  All patients of Pastorello et al. (1994) showed a positive SPT.

  SPT responses with rPru av 1, rPru av 4, and rPru av 3 were positive in 92%, 17%, and 4% respectively of the patients in group 1; in 74%, 30%, and 0% respectively of the patients in group 2; in 0%, 22%, and 89% respectively of the patients in group 4; and negative for all nonatopic subjects (group 3). Thus the sensitivity of a positive SPT response to at least one of the 3 RAs was 96%. The specificities, negative predictive values, and positive predictive values with the 3 RAs were 100%, 96%, and 100% if calculated in relation to the nonatopic control group but 17%, 79%, and 60% when calculated in relation to the control group with birch pollen allergy (Ballmer-Weber et al. 2002)

IgE assay (by RAST, CAP etc)

• Number of Studies:0
• Food/Type of allergen:

  Cherry extract and recombinant cherry Pru av 1. Fresh fruits were cut and homogenised in acetone/dry ice at -60 to -65°C ant the mixture was stored overnight and cooled with dry ice. The resulting precipitates were washed twice with acetone (–60°C) and once with chilled acetone-diethyl ether (1:1, –60°C). The filter residue was lyophilized. 2 gr of the obtained acetone powder was extracted with 30 mL of PBS (0.01M potassium phosphate and 0.15 M NaCl (pH 7.4)). The suspension was centrifuged for 45 minutes at 20,000g, and the supernatant was filtered through a fiber glass prefilter (Scheurer et al. 1997)

  Cherry extract (prepared as for Scheurer et al. 1997) and recombinant cherry LTP, Pru av 1 and Pru av 4 (Scheurer et al. 2001)

• IgE protocol:

  EAST (Scheurer et al. 1997 and 2001)

  Histamine release (Scheurer et al. 1997)

• Number of Patients:

  19 patients allergic to birch pollen and cherries (Scheurer et al. 1997).

  101 German patients with a clinical history of OAS to cherry and birch pollen and 7 Italian patients with reactions in open food challenges to cherry (Scheurer et al. 2001).

• Summary of Results:

  89 % of the patients had IgE to rPru a 1 with 18 sera from 19 individuals having IgE which reacted with cherry extract. For all the patients the maximal histamine release was at a concentration of 0.01 mg/ml (Scheurer et al. 1997).

  96 % of German patients had IgE specific to rPru av 1 whilst 3% were positive to rPru av 3 (LTP) and 16% to rPru av 4. In the Italian patients 100% were IgE positive to rPru av 3, with 28% patients positive to rPru av 1 and rPru av 4 (Scheurer et al. 2001)

Immunoblotting

• Immunoblotting separation:

  The extracts were separated in a discontinuous buffer system in an SDS-polyacrylamide gradient gel with a 6% stacking gel and a 7.5% to 20% separation gradient. Samples were boiled and reduced with beta-mercaptoethanol (Pastorello et al. 1994) [156]

  Proteins from cherry extract were separated by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) under nonreducing conditions. Purified recombinant profilins were separated under non-reducing conditions (Scheurer et al. 1997).

  Proteins from cherry extracts were separated on 12% SDS-PAGE under reducing conditions(Inschlag et al. 1998) (Scheurer et al. 2001).

  Proteins were separated by means of SDS-PAGE by using a Bio-Rad Mini Protean cell. The 15% separating gel was overlaid with a 5% stacking gel. Recombinant Pru av 4 was reduced with 1,4-dithiothreitol and rPru av 1 and rPru av 3 were applied under nonreducing conditions (Ballmer-Weber, et al 2002).

• Immunoblotting detection method:

  The proteins were electroblotted to a nitrocellulose membrane, pore size 0.2 to 0.45 µm using a Trans-Blot Cell. The membrane was blocked with phosphate-buffered saline pH 7.4 with 0.5% (v/v) Tween 20 and incubated with the sera (diluted 1:4). The IgE-binding components were detected with iodine 125–labeled anti-human IgE antiserum diluted 1:4 (Pastorello et el. 1994) [156].

  Separated proteins were electroblotted onto nitrocellulose membrane pore size 0.45 µm. The membrane was blocked with phosphate-buffered saline pH 7.4 with 0.3% (v/v) Tween 20 and incubated with the sera (diluted 1:6). The IgE-binding components were detected with peroxidase conjugated anti-human IgE antiserum and developed with ECL Western blotting detection reagents (Scheurer et al. 1997)

  Separated proteins were electroblotted onto nitrocellulose membrane pore size 0.45 µm. The membrane was incubated with the cherry allergic patients and the IgE-binding components were detected with iodine 125–labeled anti-human IgE rabbit antibody (Inschlag et al. 1998)

  Protein was electroblotted onto nitrocellulose membrane pore size 0.45 µm. The membrane was incubated with the sera (diluted 1:6.7) and the IgE-binding was detectec with an alkaline phosphatase labelled mouse anti-human IgE antibody. Antibody binding was visualised by the AP conjugate substrated kit (BioRad) (Scheurer et al. 2001)

  The proteins were transferred onto 0.2 µm of nitrocellulose membranes by means of tank blotting. The nitrocellulose membrane was blocked and incubated with patient sera. IgE antibody detection was performed with alkaline phosphatase-conjugated mouse anti-human IgE (1:750) and blots were developed with the AP Conjugate Substrate Kit (Ballmer-Weber et al. 2002).

• Immunoblotting results:

  88% of sera (16 of 18) of Pastorello et al. 1994 [156], showed Ig E binding to a 13 kDa protein and 77% (14 of 18) to a 30 kDa protein . The other IgE -binding components were: 20 kDa (8 of 18, 44%), 70 kDa (7 of 18, 38%), 14 kDa (6 of 18, 33%), 48 kDa (5 of 18, 27%), 17 kDa (3 of 18, 16%), and 50 kDa (3 of 18, 16%). Of the seven patients with negative responses to birch pollen, five showed only one IgE-binding component at 13 kDa, and two (patients 5 and 6) also had a second band at 30 kDa.

  IgE from all tested sera (6/6) reacted with a dominant band of 18-19 kDa in the cherry extract. Additional bands of lower intensity (15 kDa) were recognised by four sera and of higher molecular masses (34, 55, 65 kDa) by two sera (Scheurer et al. 1997).

  All sera tested (5) displayed IgE binding to the 30 kDa component. Three patients also reacted to proteins at 67-90 kDa and one of them to a 17, 32 and 40 kda proteins (Inschlag et al. 1998)

  All patients with allergy to cherry (13/49) showed reactivity to the cherry profilin (Scheurer et al. 2001).

  92% of sera of Ballmer-Weber et al. 2002 showed IgE binding to cherry extract on immunoblots. All patients' sera positive to one of the recombinant allergens reacted with polypeptides in the molecular weight range between 9 kDa and approximately 18 kDa, corresponding to Pru av 1, Pru av 3, and Pru av 4. In addition, weakly IgE reactivite polypeptides were observed in the higher molecular weight range (>50 kDa), which probably represents proteins with cross-reactive carbohydrate determinants. No IgE reactivity was found against a putative 23-kd allergen, which has been described as a thaumatin-like protein.

Oral provocation

• Number of Studies:1-5
• Food used and oral provocation vehicle:

  Cherry (Pastorello et al. 1994) [156] and/or blind (Ballmer-Webner et al. 2002).

  DBPCFCs were performed by means of a 2-step spit (local mucosal challenge) and swallow procedure. Two different drinks, identical in color, texture, and taste, were prepared contained 75 g of pitted cherries, 15 g of wheat flour, 10 g of cabbage, 6 teaspoons of mint syrup, 15 g of cocoa, a pinch of saffron, and 135 g of water mixed in a blender. The placebo drink contained the same ingredients but no cherries and, in addition, 13 g of sugar and 27 g of beetroot juice (Ballmer-Weber et al. 2002).

  In patients with birch pollen allergy but without cherry allergy and nonatopic control subjects, an open challenge was performed. These patients had to chew and swallow 6 fresh cherries (Ballmer-Weber et al. 2002)

• Blind:Either open or blind challenges.
• Number of Patients:

  21 patients (Pastorello et al. 1994) [156]

  79 subjects were included in the study: 24 Swiss patients (group 1) with a positive double-blind placebo-controlled food challenge result to cherries, 23 patients with birch pollen allergy but without a history of cherry allergy (group 2), 23 nonatopic subjects (group 3), and 9 Spanish patients with a history of a cherry allergy (group 4) (Ballmer-Weber et al. 2002).

• Dose response:

  Patients chewed the fruit for 1 minute and then spit it out. If no symptoms appeared within 15 minutes, the challenge was repeated with increasing amounts from 250mg up to 64 g. Patients were asked not to spit out the last two doses but to swallow the fruit instead. 60 g of cherry fruit was required to elicit a reaction (Pastorello et al. 1994) [156]
  

  3.3 g of cherries (Ballmer-Weber et al. 2002).

• Symptoms:

  Oropharyngeal sympotoms (itching or tingling of the lips or oral mucosa) in 8 patients and gastrointestinal symptoms (abdominal pain, vomiting) in 2 patients (Pastorello et al. 1994) [156]

  In the DBPCFCs 16 patients complained about symptoms strictly localized to the oral cavity (oral allergy syndrome, OAS). In 7 patients, symptoms were not restricted to the oral cavity but also dyspnea, dysphagia, rhinitis, gastrointestinal symptoms. One patient was not challenged because she had a severe allergic reaction with urticaria, angioedema, severe dyspnea, cough, and gastrointestinal symptoms accompanied by severe pain after cherry consumption. Subjects with birch allergy and no history of cherry allergy (group 2) and non-atopic (group 3) did not experience any reaction during open provocation with fresh cherries (Ballmer-Weber et al. 2002).

IgE cross-reactivity and Polysensitisation

Other Clinical information

Reviews (0)

References (6)

• Ballmer-Weber BK, Scheurer S, Fritsche P, Enrique E, Cistero-Bahima A, Haaseb T, Wüthrich B
  Component-resolved diagnosis with recombinant allergens in patients with cherry allergy
  J. Allergy Clin. Immunol. 110 (1). 2002
  PUBMEDID: 12110837
• Scheurer S, Wangorsch A, Nerkamp J, Skov PS, Ballmer-Weber B, Wuthrich B, Haustein D, Vieths S.
  Cross-reactivity within the profilin panallergen family investigated by comparison of recombinant profilins from pear (Pyr c 4), cherry (Pru av 4) and celery (Api g 4) with birch pollen profilin Bet v 2.
  J Chromatogr B Biomed Sci Appl May 756:315-325.. 2001
  PUBMEDID: 11419723
• Scheurer S, Metzner K, Haustein D, Vieths S.
  Molecular cloning, expression and characterisation of Pru a 1, the major cherrry allergen.
  Mol. Immunol. 34:619-629.. 1997
  PUBMEDID: 9393965
• Pastorello EA, Ortolani C, Farioli L, Pravettoni V, Ispano M, Borga A, Bengtsson A, Incorvaia C, Berti C, Zanussi C.
  Allergenic cross-reactivity among peach, apricot, plum, and cherry in patients with oral allergy syndrome: an in vivo and in vitro study.
  J Allergy Clin Immunol 94(4):699-707. 1994
  PUBMEDID: 7930303
• Scheurer S, Pastorello EA, Wangorsch A, Kastner M, Haustein D, Vieths S.
  Recombinant allergens Pru av 1 and Pru av 4 and a newly identified lipid transfer protein in the in vitro diagnosis of cherry allergy.
  J Allergy Clin Immunol 107:724-31.. 2001
  PUBMEDID: 11295665
• Inschlag C, Hoffmann-Sommergruber K, O'Riordain G, Ahorn H, Ebner C, Scheiner O, Breiteneder H.
  Biochemical characterisation of Pru a 2, a 23kD thaumatin-like protein representing a potential major allergen in cherry (Prunus avium)
  Int. Arch. Allergy Immunol. 116:22-28.. 1998
  PUBMEDID: 9623505

Biochemical Information for Pru av 1

• Allergen Name:Pru av 1
• Alternatve Allergen Names:Previously known as Pru a 1 but now renamed
• Allergen Designation:Major
• Protein Family:

  Pathogenesis-related protein Bet v I family, Pfam, PF00407

• Sequence Known?:Yes
• Allergen accession No.s:

  O24248:Swissprot:http://ca.expasy.org/cgi-bin/niceprot.pl?O24248

  U66076; AAC02632.1; -. EMBL

  Q6QHU3:Swissprot:http://us.expasy.org/cgi-bin/niceprot.pl?Q6QHU3

  AY540507; AAS47035.1; -.

  EMBL

  Q6QHU2:Swissprot:http://us.expasy.org/cgi-bin/niceprot.pl?Q6QHU2

  AY540508; AAS47036.1; -.

  EMBL

  Q6QHU1:Swissprot:http://us.expasy.org/cgi-bin/niceprot.pl?Q6QHU1

  AY540509; AAS47037.1; -.

  EMBL

• 3D Structure Accession No.:

  1E09 http://www.rcsb.org/pdb/cgi/explore.cgi?pdbId=1E09

• Calculated Masses:

  17 660, 17 351, 17 340, 17 342 Da

• Experimental Masses:18 kDa
• Oligomeric Masses:
• Allergen epitopes:

  Five overlapping recombinant Pru av 1 fragments representing the entire amino acid sequence corresponding to the 'P-loop' region, with lengths of approximately 60-120 residues have been investigated. Weak IgE binding capacity was measured exclusively with Pru av IF4 (residues 1-120) by immunoblotting, but none of the fragments showed allergenicity in the rat basophil leukaemia cell mediator release assay. Site-directed mutagenesis experiments with Pru av 1 revealed that amino acid S112 is critical for IgE binding of almost all patients sera tested. This residue appears to be essential residue for preserving the structure of a cross-reactive IgE epitope. (Scheurer et al 1999).

  Mutagenesis of Glu⁴⁵ and Ser¹¹² of Pru av 1 to Pru av 1 Trp⁴⁵, Ala¹¹², Pro¹¹² was performed by PCR and the IgE reactivity determined by EAST and immunoblotting. Disruption of the native tertiary structure of Pru av 1 upon the mutation of Pro¹¹² leads to the nearly complete loss of the IgE reactivity indicating that the cross-reactive IgE-binding epitopes of Pru av 1 are predominantly conformational rather than sequential. The mutation of Glu⁴⁵ to Trp⁴⁵ in the P-loop region, significantly reduced IgE binding to Pru av 1 in a subgroup of cherry-allergic patients therefore, this residue is involved in a cross-reactive IgE epitope. The mutation Ala¹¹² did not influence IgE binding to Pru av 1 (Neudecker et al. 2003)

• Allergen stability:
  Process, chemical, enzymatic:

  Strong thermolability was shown by CD-spectroscopy (loss of secondary structure at 70°C) and by IgE-reactivity of thermal processed food. Lability to enzymatic digestion was shown by pepsin treatment of cherry extract and recombinant Pru av 1 (stability: Pru av 3 > Pru av 1> Pru av 4) (Scheurer et al. 2004).

  Influence of thermal processing and nonenzymatic as well as enzymatic browning reactions on Pru av 1 was investigated by Gruber et al (2004). Thermal unfolding of the allergen occurred at a calculated mellting temperature of 66.3°C. Correct refolding occurred after cooling. Allergenicity of Pru av 1 was reduced by enzymatic polyphenol oxidation and Maillard reaction induced by carbohydrate breakdown products.

• Nature of main cross-reacting proteins:

  As a consequnece of high sequence identity to other members of the Bet v 1 family IgE to Pru av 1 cross-reacts with pathogenesis related proteins from parsley, potato, soya bean, Mal d 1 from apple, and Bet v 1 from birch pollen (Scheurer et al. 1997, 1999).

• Allergen properties & biological function:Pru av 1 is a member of pathogenesis-related protein group 10. These proteins are expressed upon pathogen attack, stress and abiotic stimuli. Although, Pru av 1 precise function is not known, same evidence is given that Pru av1 is involved in steroid binding (Neudecker et al 2000; Neudecker et al 2001).
• Allergen purification:

  Recombinant Pru av 1has been purified using a heterologous expression system, E. coli, as a His tag fusion portein and further purified by metal chelate affinity chromatography (Scheurer et al. 1997).

• Other biochemical information:

  Severeal isoforms have been identified. Pru av 1.0101, Pru av 1.0201, Pru av 1.0202, Pru av 1.0203 (Scheurer et al. 1997) [176]; (Reuter et al. 2004) [1163]

References (9)

• Neudecker P, Lehmann K, Nerkamp J, Haase T, Wangorsch A, Fotisch K, Hoffmann S, Rosch P, Vieths S, Scheurer S
  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
  Biochem. J. 376: 97-107 Part 1 NOV 15. 2003
  PUBMEDID: 12943529
• Scheurer S, Son DY, Boehm M, Karamloo F, Franke S, Hoffmann A, Haustein D, Vieths S
  Cross-reactivity and epitope analysis of Pru a 1, the major cherry allergen.
  Mol Immunol 36:155-67.. 1999
  PUBMEDID: 10403481
• Neudecker P, Schweimer K, Nerkamp J, Scheurer S, Vieths S, Sticht H, Rosch P.
  Allergic Cross-reactivity Made Visible. Solution structure of the major cherry allergen Pru av 1.
  J Biol Chem 276:22756-63.. 2001
  PUBMEDID: 11287426
• Neudecker P, Schweimer K, Nerkamp J, Boehm M, Scheurer S, Vieths S, Sticht H, Rosch P
  Sequence-specific 1H, 13C and 15N resonance assignments of the major cherry allergen Pru a 1.
  J Biomol NMR 18:71-2.. 2000
  PUBMEDID: 11061231
• Scheurer S, Pastorello EA, Wangorsch A, Kastner M, Haustein D, Vieths S.
  Recombinant allergens Pru av 1 and Pru av 4 and a newly identified lipid transfer protein in the in vitro diagnosis of cherry allergy.
  J Allergy Clin Immunol 107:724-31.. 2001
  PUBMEDID: 11295665
• Scheurer S, Metzner K, Haustein D, Vieths S.
  Molecular cloning, expression and characterisation of Pru a 1, the major cherrry allergen.
  Mol. Immunol. 34:619-629.. 1997
  PUBMEDID: 9393965
• Gruber P, Vieths S, Wangorsch A, Nerkamp J, Hofmann T
  Maillard reaction and enzymatic browning affect the allergenicity of Pru av 1, the major allergen from cherry (Prunus avium)
  J Agric Food Chem. 52(12):4002-7. 2004
  PUBMEDID: 15186129
• Scheurer S, Lauer I, Foetisch K, Moncin MS, Retzek M, Hartz C, Enrique E, Lidholm J, Cistero-Bahima A, Vieths S
  Strong allergenicity of Pru av 3, the lipid transfer protein from cherry, is related to high stability against thermal processing and digestion
  J Allergy Clin Immunol. 114(4):900-7.. 2004
  PUBMEDID: 15480332
• Reuter A, Fortunato D, Perono Garoffo L, Napolitano L, Scheurer S, Giuffrida MG, Vieths S, Conti A
  Novel isoforms of Pru av 1 with diverging immunoglobulin E binding properties identified by a synergistic combination of molecular biology and proteomics
  Proteomics. Dec 9; [Epub ahead of print]. 2004
  PUBMEDID: 15593144

Biochemical Information for Pru av 2

• Allergen Name:Pru av 2
• Alternatve Allergen Names:
• Allergen Designation:Major
• Protein Family:Thaumatin family, Pfam, PF00314
• Sequence Known?:Yes
• Allergen accession No.s:

  P50694:Swissprot:http://ca.expasy.org/cgi-bin/niceprot.pl?P50694

  U32440; AAB38064.1; - EMBL

• 3D Structure Accession No.:Not known
• Calculated Masses:23336.03 Da
• Experimental Masses:29 kDa
• Oligomeric Masses:
• Allergen epitopes:Not known
• Allergen stability:
  Process, chemical, enzymatic:

  The thaumatin-like proteins (TLPs) contain 8 disulfide bridgesand so it might be expected to be thermostable. The TPLs are generally resistant to proteases and pH-induced denaturation (Breiteneder, H. 2004) [1001]

• Nature of main cross-reacting proteins:Not known
• Allergen properties & biological function:Pru av 2 has significant similarities to other thaumatin-like proteins belonging to pathogenesis-related protein group 5 and are thought to be produced in response to pathogen infection or to osmotic stress. (Inschlag et al. 1998)
• Allergen purification:

  Pru av 2 was purified using 0.2M sodium acetate, 1.4M sodium chloride extraction buffer containing PVP. A precipitation step followed using 80 % ammonium sulfate. The proteins were chromatographed by size exclusion (ACA 54, 100x1.6 cm) and eluted with 0.1M sodium acetate, and anion exchange (DEAE-Sepharose CL6B, 10x2 cm) and eluted with 0.03M sodium acetate (Fils-Lycaon et al. 1996)

  Pru av 2 was purified by reverse phase HPLC on a C8 (Hypersil WP300,, 8x250mm, 10um particle size) using 0.1% (v:v) trifluroacetic acid as the solvent and eluting the TLP with a linear gradient of propan-2-ol (0-80%, 60min, 1.0ml/min). (Insclag et al 1998).

• Other biochemical information:

  Pru av 2 has a pI of 4.2 (Fils-Lycaon et al. 1996).

References (4)

• Fils-Lycaon B.R., Wiersma P.A., Eastwell K.C., Sautiere P.
  A cherry protein and its gene, abundantly expressed in ripening fruit, have been identified as thaumatin-like.
  Plant Physiol. 111:269-273. 1996
  PUBMEDID: 8685266
• Pastorello EA, Farioli L et al
  Identification of a major allergen from cherry as a thaumatin-like protein
  Giorn It Allergol Immunol :24-29. 1999
  PUBMEDID:
• Inschlag C, Hoffmann-Sommergruber K, O'Riordain G, Ahorn H, Ebner C, Scheiner O, Breiteneder H.
  Biochemical characterisation of Pru a 2, a 23kD thaumatin-like protein representing a potential major allergen in cherry (Prunus avium)
  Int. Arch. Allergy Immunol. 116:22-28.. 1998
  PUBMEDID: 9623505
• Breiteneder H.
  Thaumatin-like proteins -- a new family of pollen and fruit allergens
  Allergy. 59:79-81.. 2004
  PUBMEDID: 15080826

Biochemical Information for Pru av 3

• Allergen Name:Pru av 3
• Alternatve Allergen Names:non specific lipid transfer protein (nsLTP)
• Allergen Designation:Minor (Central European population) and major (Mediterranean population).
• Protein Family:Protease inhibitor/seed storage/LTP family, Pfam PF00234
• Sequence Known?:Yes
• Allergen accession No.s:

  Q9M5X8:Swissprot: http://ca.expasy.org/cgi-bin/niceprot.pl?Q9M5X8

  AF221501; AAF26449.1; -. EMBL

• 3D Structure Accession No.:Not known
• Calculated Masses:9147.54 Da
• Experimental Masses:9.2 kDa
• Oligomeric Masses:
• Allergen epitopes:Not known
• Allergen stability:
  Process, chemical, enzymatic:Strong thermostability was shown by CD-spectoscopy (preserved secondary structure at 70°C) and by IgE-reactivity of thermal processed food. Strong stability to enzymatic digestion was shown by pepsin treatment of cherry extract and recombinant Pru av 3 (stability: Pru av 3 > Pru av 1> Pru av 4) (Scheurer et al. 2004)
• Nature of main cross-reacting proteins:Sequence identity of Pru av 3 with that of almond (89%), peach Pru p 3 (87.9%) and apricot Pru ar 3 (85.7%) indicates that IgE cross-reactivity between these proteins is likely.
• Allergen properties & biological function:

  Plant nonspecific lipid-transfer proteins are thought to be involved in transport of fatty acids both intracellular and extracellularly and of cutin monomers to the cuticular layer of leaves and fruits. There is an expandable cavity between the four alpha-helices which can bind one or two lipids. nsLTPs have also been reported to act as plant defense proteins against bacterial and fungal infections and form the PR14 family of pathogenesis related proteins. It is possible that a lipid-like post-translational modification is involved (Lindorff-Larsen et al. 2001 [903]).
  

• Allergen purification:Cherry LTP purified from fruit has not been reported, but prepared using a heterologous expression system, E. coli, as a His tag fusion protein and was purified on nickel-nitrilotriacetate solid-phase (Scheurer et al. 2001)
• Other biochemical information:

  Pru av 3 has a pI of 9.22.

References (3)

• Scheurer S, Pastorello EA, Wangorsch A, Kastner M, Haustein D, Vieths S.
  Recombinant allergens Pru av 1 and Pru av 4 and a newly identified lipid transfer protein in the in vitro diagnosis of cherry allergy.
  J Allergy Clin Immunol 107:724-31.. 2001
  PUBMEDID: 11295665
• Lindorff-Larsen K, Lerche MH, Poulsen FM, Roepstorff P, Winther JR.
  Barley lipid transfer protein, LTP1, contains a new type of lipid-like post-translational modification.
  J Biol Chem. 276(36):33547-33553.. 2001
  PUBMEDID: 11435437
• Scheurer S, Lauer I, Foetisch K, Moncin MS, Retzek M, Hartz C, Enrique E, Lidholm J, Cistero-Bahima A, Vieths S
  Strong allergenicity of Pru av 3, the lipid transfer protein from cherry, is related to high stability against thermal processing and digestion
  J Allergy Clin Immunol. 114(4):900-7.. 2004
  PUBMEDID: 15480332

Biochemical Information for Pru av 4

• Allergen Name:Pru av 4
• Alternatve Allergen Names:Has previously been described as Pru a 3 before being renamed
• Allergen Designation:Minor
• Protein Family:Profilin, Pfam, PF00235
• Sequence Known?:Yes
• Allergen accession No.s:

  Q9XF39:Swissprot:http://ca.expasy.org/cgi-bin/niceprot.pl?Q9XF39

  AF129425; AAD29411.1; -. EMBL

• 3D Structure Accession No.:Not known
• Calculated Masses:13970 Da
• Experimental Masses:14 kDa
• Oligomeric Masses:
• Allergen epitopes:Not known
• Allergen stability:
  Process, chemical, enzymatic:

  There are only few studies on the stability of profilins most of them on celery profilin. Compared to other allergens, profilin is a moderately stable protein, more resistant than Bet v 1 homologues but less stable than lipid transfer proteins or cross-reactive carbohydrate deteminants of glycoprotein allergens.

  High lability to enzymatic digestion was shown by pepsin treatment of cherry extract and recombinant Pru av 4 (stability: Pru av 3 > Pru av 1> Pru av 4). (Scheurer et al. 2004)
• Nature of main cross-reacting proteins:As consequence of high sequence identities, IgE to Pru av 4 cross-reacts with the profilin from birch pollen Bet v 2 (76.1%), from celery Api g 4 (76.9%) and from tomato Lyc e 1(Scheurer et al 2001) (Westpahl et al. 2004)
• Allergen properties & biological function:

  Pru av 4 is thought to be an actin-binding protein of the cytoskeleton

• Allergen purification:Recombinant Pru av 4 has been purified using a heterologous expression system, E. coli, and further purified on PLP-coupled sepharose.
• Other biochemical information:

  Pru av 4 has a pI of 4.47

References (4)

• Scheurer S, Wangorsch A, Nerkamp J, Skov PS, Ballmer-Weber B, Wuthrich B, Haustein D, Vieths S.
  Cross-reactivity within the profilin panallergen family investigated by comparison of recombinant profilins from pear (Pyr c 4), cherry (Pru av 4) and celery (Api g 4) with birch pollen profilin Bet v 2.
  J Chromatogr B Biomed Sci Appl May 756:315-325.. 2001
  PUBMEDID: 11419723
• Scheurer S, Pastorello EA, Wangorsch A, Kastner M, Haustein D, Vieths S.
  Recombinant allergens Pru av 1 and Pru av 4 and a newly identified lipid transfer protein in the in vitro diagnosis of cherry allergy.
  J Allergy Clin Immunol 107:724-31.. 2001
  PUBMEDID: 11295665
• Scheurer S, Lauer I, Foetisch K, Moncin MS, Retzek M, Hartz C, Enrique E, Lidholm J, Cistero-Bahima A, Vieths S
  Strong allergenicity of Pru av 3, the lipid transfer protein from cherry, is related to high stability against thermal processing and digestion
  J Allergy Clin Immunol. 114(4):900-7.. 2004
  PUBMEDID: 15480332
• Westphal S, Kempf W, Foetisch K, Retzek M, Vieths S, Scheurer S
  Tomato profilin Lyc e 1: IgE cross-reactivity and allergenic potency
  Allergy. 59(5):526-32. 2004
  PUBMEDID: 15080834