Breiteneder H, Ferreira F, Reikerstorfer A, Duchene M, Valenta R, Hoffmann-Sommergruber K, Ebner C, Breitenbach M, Kraft D, Scheiner O
Institute of General and Experimental Pathology, University of Vienna, Austria.
J Allergy Clin Immunol. 1992 Dec;90(6 Pt 1):909-17. doi: 10.1016/0091-6749(92)90463-c.
Previous data showed that the major pollen allergens from trees of the order Fagales, in particular alder, birch, hazel, and hornbeam, are highly interrelated. As only the complete amino acid sequence of Bet v I, the major allergen from birch, has been known, it was of interest to obtain the primary structure of other major allergens of this group, to attribute IgE-binding properties to certain features of the amino acid sequences of those allergens. cDNA was synthesized from alder pollen mRNA, sequence-specifically amplified by polymerase chain reaction and cloned into plasmid bluescript. Comparison of the deduced amino acid sequences of Aln g I and Bet v I revealed a 86.8% homology. The Aln g I encoding cDNA was subcloned into pKK223-3 and expressed in Escherichia coli as a full-length nonfusion protein. The recombinant Aln g I bound IgE from tree pollen-allergic patients and was shown to share IgE-epitopes with Bet v I by inhibition studies with recombinant Bet v I. Computer-aided calculations predicted epitopes in both Aln g I and Bet v I at the same position; the Bet v I molecule was predicted to possess two additional epitopes near the N-terminus of the molecule.
先前的数据表明，壳斗目树木的主要花粉过敏原，特别是桤木、桦树、榛树和鹅耳枥的花粉过敏原，具有高度的相关性。由于仅知道桦树主要过敏原Bet v I的完整氨基酸序列，因此获得该组其他主要过敏原的一级结构，并将IgE结合特性归因于这些过敏原氨基酸序列的某些特征，是很有意义的。从桤木花粉mRNA合成cDNA，通过聚合酶链反应进行序列特异性扩增，并克隆到质粒pBluescript中。对Aln g I和Bet v I推导的氨基酸序列进行比较，发现同源性为86.8%。将编码Aln g I的cDNA亚克隆到pKK223-3中，并在大肠杆菌中表达为全长非融合蛋白。重组Aln g I与树花粉过敏患者的IgE结合，并通过重组Bet v I的抑制研究表明与Bet v I共享IgE表位。计算机辅助计算预测Aln g I和Bet v I在相同位置具有表位；预测Bet v I分子在分子N端附近还具有两个额外的表位。
