Immunological and biological properties of Bet v 4, a novel birch pollen allergen with two EF-hand calcium-binding domains

被引:117
作者
Engel, E
Richter, K
Obermeyer, G
Briza, P
Kungl, AJ
Simon, B
Auer, M
Ebner, C
Rheinberger, HJ
Breitenbach, M
Ferreira, F
机构
[1] SALZBURG UNIV,INST GENET & ALLGEMEINE BIOL,A-5020 SALZBURG,AUSTRIA
[2] SALZBURG UNIV,INST PFLANZENPHYSIOL,A-5020 SALZBURG,AUSTRIA
[3] NOVARTIS FORSCHUNGSINST,A-1235 VIENNA,AUSTRIA
[4] INST ALLGEMEINE & EXPT PATHOL,A-1090 VIENNA,AUSTRIA
关键词
D O I
10.1074/jbc.272.45.28630
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have isolated a cDNA clone coding for a birch pollen allergen, Bet v 4. The deduced amino acid sequence of Bet v 4 contained two typical EF-hand calcium-binding domains, Sequence similarities of Bet v 4 to calmodulin are primarily confined to the calcium-binding domains, However, significant sequence similarities extending outside the Ca2+-binding sites were found with a recently described group of pollen-specific allergens of Brassica and Bermuda grass, Both EF-hand domains of Bet v 4 are able to bind Ca2+, as demonstrated by Ca-45(2+) blot overlay of wild type and calcium-binding deficient mutants of Bet v 4, Among pollen-allergic patients, protein-bound Ca2+ was not an absolute requirement for IgE recognition of Bet v 4, However, disruption of the carboxyl-terminal Ca2+-binding domain indicated that most IgE antibodies from allergic patients are directed against this site. IgE inhibition experiments suggested that Bet v 4 represents a high;ly cross-reactive pollen allergen. Pre-absorption of allergic sera with Bet v 4 drastically reduced IgE binding to proteins of similar molecular weight in pollen extracts from distantly related plant species (e.g. timothy grass, mugwort, lily) but not in extracts from plant-derived foodstuff, To test for a possible biological role in pollen germination and tube growth, we introduced recombinant Bet v 4 protein into growing lily pollen tubes by iontophoresis. As a result, cytoplasmic streaming stopped in the vicinity of the electrode tip, and a slight depolarization of the membrane voltage was measured. These effects were not observed with Ca2+-binding deficient mutants of Bet v 4, Thus, Bet v 4 and homologous proteins represent a new class of pollen-specific Ca2+-binding allergens that may have a physiological role as inhibitors of cytoplasmic streaming in outgrowing pollen tubes.
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页码:28630 / 28637
页数:8
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