STUDY ON THE SUGAR MOIETY AS A COMMON ANTIGEN OF WHEAT ALLERGENS

小麦过敏原常见抗原糖部分的研究

• 批准号: 15580112
• 负责人: TSUJI Hideaki
• 金额: $ 2.37万
• 依托单位: Okayama Prefectural University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15580112/
• 关键词: Wheat allergen; Tri a Bd 27K; Tri a Bd 17K; Sugar chain; Allergenicity; Common antigen; Tri a Bd17K; Tri a Bd 36K; common epitope

Many plant allergens have been found, and show more complex phase concerning their allergen investigation field. Crossreaction between the allergens have been very often observed. Therefore, the complex phase would be unified by the idea entitle "a common epitope". Asn-containing epitopes on the allergens is considering as one of common antigens.We have elucidated the sugar moiety of soybean major allergen, Gly m Bd 28K and wheat major allergen Tri a Bd 17K.In the present study, the properties of Tria Bd 27K, a major allergen, in wheat, has been investigated in details. First of all, monoclonal antibodies against the allergen have been producwed. The allergen in wheat was separated by two-dimensional electrophoresis with isoelectric electrophoresis and SDS-PAGE, and the proteins on the gel were detected by immnoblotting with the above-mentioned antibodies. Several spots showing the positive response for the antibodies were observed. The regions on the gel corresponding to the positive spots were cut out and subjected to N-terminal amino acid sequenceing. The proteins on the regions were shown to belong to the Tri a Bd 27k. This finding means that Tri a Bd 27K occur as polyforms. The above-mentioned spots reacted with a lectin, sensitive to fukose residue and the sera of patients positive to wheat. Furthermore, the Tri a Bd 27K sensitive to the lectin were cut out and subjected to sugar analyses. Subsequently, mannose, N-acetylglucosamin were detected, and the occurrence of galactose and fukose and xylose were alsoshowed. Interestingly, a recombinant Tri a Bd 27K expressed in Eschericia coli was not shown to be insetive to the sera of patients with wheat allergy.The present findings demonstrate that most of IgE antibodies in the sera of the above patients, which were sensitive to the allergen, bind to the sugar moiety of the allergen.

已经发现了许多植物过敏原，并显示出有关其过敏原研究场的更复杂的阶段。经常观察到过敏原之间的交叉反应。因此，复杂阶段将由“共同表位”权利的想法统一。过敏原上的含有的表位被认为是一种常见的抗原。我们阐明了大豆主要过敏原的糖部分，Gly M BD 28K和小麦主要的过敏原Tri 17K。首先，对过敏原的单克隆抗体进行了生产。小麦中的过敏原通过二维电泳与等电电泳和SDS-PAGE分离，并通过与上述抗体进行INMNOBROTIES检测凝胶上的蛋白质。观察到几个斑点显示了对抗体的阳性反应。切出与阳性斑点相对应的凝胶区域，并进行N端氨基酸序列。该区域上的蛋白质属于Tri A BD 27K。这一发现意味着Tri A BD 27K以多形式出现。上述斑点与凝集素反应，对富酮残基敏感和阳性小麦的血清。此外，将对凝集素敏感的Tri A BD 27K被切除并进行糖分析。随后，检测到甘露糖，N-乙酰葡萄糖，并将半乳糖和福酮和木糖的发生。有趣的是，在大肠杆菌中表达的重组TRI A BD 27K并未证明对小麦过敏患者的血清具有固定性。目前的发现表明，上述患者血清中的大多数IgE抗体对过敏原敏感，与过敏原敏感。

项目成果

Identification of the 23-kDa peptide derived from th precursor of Gly m Bd 28K, a major soybean allergen, as a new allergen.

将源自主要大豆过敏原 Gly m Bd 28K 前体的 23-kDa 肽鉴定为新过敏原。

• 发表时间: 2004
• 期刊: Biochim.Biophys.Acta. 1673(1-2)
• 作者: M.Hiemori;et al.
• 通讯作者: et al.