Department of Internal Medicine, Institute of Allergy, Yonsei University College of Medicine, Seoul 03722, Korea.
Protein Pept Lett. 2021;28(4):362-371. doi: 10.2174/0929866527666200813201924.
Japanese hop is an important cause of weed pollinosis in East Asia. Its pollen is abundant in autumn. This pollen is known to be the cause of many allergic diseases. However, molecular characteristics of its allergens have not been elucidated.
In this study, we produced recombinant proteins of allergen homologues from Japanese hop by the analysis of expressed sequence tags (EST), and evaluated its allergenicity.
cDNA library was constructed using as little as 50 ng of total RNA from Japanese hop pollen. Allergen homologues were identified by the initial screening of 963 EST clones. Recombinant proteins were overexpressed in the E. coli expression system and purified using Ni-nitrilotriacetic acid-agarose. Purified proteins were analyzed by ELISA.
Japanese hop pathogenesis-related 1 protein (PR-1) shares 37.0 to 44.4% of amino acid sequence identity with Art v 2, Cuc m 3, and Cyn d 24. Pectin methyl esterase (PME) shows 23.2 to 50.2% of identities to Act d 7, Ole e 11, and Sal k 1. Polygalacturonase (PGs) shows 16.7 to 19.3% of identities to Phl p 13, Cry j 2, Cha o 2, Jun a 2, Pla a 2, and Pla or 2. IgE antibodies from Japanese hop allergy patients' sera recognized PR-1 (3.4%), PME (13.8%), PGs (3.7%), and profilin (13.8%), respectively.
Novel allergenic components were identified, even though low IgE reactivity was displayed reflecting the low degree of cross-reactivity with other pollen allergens. We believe that these molecules have worth further studies.
葎草是东亚杂草花粉过敏的重要原因。它的花粉在秋季丰富。这种花粉是许多过敏性疾病的已知原因。然而，其过敏原的分子特征尚未阐明。
在这项研究中，我们通过分析表达序列标签（EST），从葎草花粉中产生了过敏原同源物的重组蛋白，并评估了其变应原性。
使用来自日本葎草花粉的总 RNA 少至 50ng 构建 cDNA 文库。通过对 963 个 EST 克隆的初步筛选鉴定过敏原同源物。在大肠杆菌表达系统中过表达重组蛋白，并使用 Ni-亚氨基三乙酸琼脂糖纯化。通过 ELISA 分析纯化的蛋白质。
日本葎草发病相关 1 蛋白（PR-1）与 Art v 2、Cuc m 3 和 Cyn d 24 的氨基酸序列同源性为 37.0 至 44.4%。果胶甲酯酶（PME）与 Act d 7、Ole e 11 和 Sal k 1 的同源性为 23.2 至 50.2%。多聚半乳糖醛酸酶（PGs）与 Phl p 13、Cry j 2、Cha o 2、Jun a 2、Pla a 2 和 Pla or 2 的同源性为 16.7 至 19.3%。日本葎草过敏患者血清中的 IgE 抗体分别识别 PR-1（3.4%）、PME（13.8%）、PGs（3.7%）和丝氨酸蛋白酶抑制剂（13.8%）。
即使与其他花粉过敏原的交叉反应性低，也鉴定出了新型变应原成分，反映出低 IgE 反应性。我们相信这些分子值得进一步研究。
