Phenylcoumaran benzylic ether and isoflavonoid reductases are a new class of cross-reactive allergens in birch pollen, fruits and vegetables.
We investigated the biochemical function of the birch pollen allergen Bet v 6 and its role in the IgE-cross-reactivity between birch pollen and plant foods, and characterized Pyr c 5, a Bet v 6-related food allergen, from pear; the proteins were expressed as His-Tag fusion proteins in Eschershia coli and purified by Ni-chelate affinity chromatography under native conditions. Nonfusion proteins were obtained by factor Xa protease treatment. The highest degree of amino-acid sequence identity of Pyr c 5 and Bet v 6 was found with a plant protein related to a defense mechanism, which we have named phenylcoumaran benzylic ether reductase (PCBER) based on its ability to catalyze the NADPH-dependent reduction of 8-5' linked lignans such as dehydrodiconiferyl alcohol to give isodihydrodehydrodiconiferyl alcohol. Enzymatic assays with recombinant Pyr c 5 and Bet v 6 showed PCBER catalytic activity for both recombinant allergens. Both Pyr c 5 and Bet v 6 allergens had similar IgE binding characteristics in immunoblotting and enzyme allergosorbent tests (EAST), and bound IgE from 10 sera of birch-pollen-allergic patients including six pear-allergic subjects. EAST inhibition experiments with Pyr c 5 as the solid phase antigen suggested that homologous allergens may be present in many vegetable foods such as apple, peach, orange, lychee fruit, strawberry, persimmon, zucchini (courgette), and carrot. In extracts of pear, apple, orange, and persimmon, the presence of proteins of approximately 30-35 kDa containing Bet v 6 cross-reactive epitopes was demonstrated with two Bet v 6-specific monoclonal antibodies. Recombinant Pyr c 5 triggered a strong, dose-dependent mediator release from basophils of a pear-allergic subject, suggesting that Pyr c 5 has the potential to elicit type I allergic reactions.
Karamloo
F
F
Swiss Institute of Allergy and Asthma Research, Davos Platz, Switzerland.
Wangorsch
A
A
Kasahara
H
H
Davin
L B
LB
Haustein
D
D
Lewis
N G
NG
Vieths
S
S
D000485:Allergens / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000595:Amino Acid Sequence; D029662:Betula / Q000201:enzymology* / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D003429:Cross Reactions / Q000276:immunology; D005512:Food Hypersensitivity / Q000276:immunology; D005638:Fruit / Q000201:enzymology / Q000276:immunology*; D006636:Histamine Release; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D007106:Immune Sera / Q000276:immunology; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000276:immunology; D008969:Molecular Sequence Data; D009249:NADP / Q000378:metabolism; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D044925:Oxidoreductases Acting on CH-CH Group Donors; D011058:Pollen / Q000201:enzymology / Q000276:immunology*; D017386:Sequence Homology, Amino Acid; D014675:Vegetables / Q000201:enzymology / Q000276:immunology*
D000485:Allergens; D007106:Immune Sera; D007073:Immunoglobulin E; D009249:NADP; D010088:Oxidoreductases; C071072:vestitone reductase; D044925:Oxidoreductases Acting on CH-CH Group Donors; C118150:phenylcoumaran benzylic ether reductase