Induction of IgE antibody responses by glutathione S-transferase from the German cockroach (Blattella germanica)
Induction of IgE antibody responses by glutathione S-transferase from the German cockroach (Blattella germanica)
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DOI：
10.1074/jbc.272.33.20907
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发表时间：
1997-08-15
影响因子：
4.8
通讯作者：
Chapman, MD
中科院分区：
文献类型：
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作者：
Arruda, LK;Vailes, LD;Chapman, MD
我们报道了德国小蠊（Blattella germanica）的主要23 kda过敏原是谷胱甘肽s -转移酶（EC 2.5.1.18； GST）。采用谷胱甘肽亲和层析法从蟑螂体提取物中纯化天然德国小蠊GST，并利用pET21a载体在大肠杆菌中表达重组蛋白，显示出良好的IgE抗体结合活性。德国小蠊GST在使用3 pg重组蛋白的蟑螂过敏患者中引起立即皮肤试验阳性。该天然蛋白的nh2末端序列与cDNA推导出的氨基酸序列除有一个取代（Phe(9)—> Cys）外完全相同。该蛋白与谷胱甘肽结合，序列与冈比亚按蚊（Anopheles gambiae）和黑腹果蝇（Drosophila melanogaster）等昆虫的GST- s亚家族同源（42-51%），被确定为GST超家族成员。通过x射线晶体学鉴定，德国芽孢杆菌GST含有哺乳动物GST中26个不变残基中的18个，并对GST底物表现出酶活性。我们的研究结果表明蟑螂GST引起IgE抗体反应，并与哮喘有关。这些数据有力地支持了对GST的免疫应答在过敏性疾病中起重要作用的观点。
We report that a major 23-kDa allergen from German cockroach (Blattella germanica) is a glutathione S-transferase (EC 2.5.1.18; GST). Natural B. germanica GST, purified from cockroach body extracts by glutathione affinity chromatography, and recombinant protein expressed in Escherichia coli using the pET21a vector, showed excellent IgE antibody binding activity. B. germanica GST caused positive immediate skin tests in cockroach-allergic patients using as little as 3 pg of recombinant protein. The NH2-terminal sequence of the natural protein and the deduced amino acid sequence from cDNA were identical except for one substitution (Phe(9) --> Cys). Assignment of this protein to the GST superfamily was based on binding to glutathione and sequence identity (42-51%) to the GST-S subfamily from insects, including Anopheles gambiae and Drosophila melanogaster. B. germanica GST contained 18 of the 26 invariable residues identified in mammalian GST by xray crystallography and exhibited enzymic activity against a GST substrate. Our results show that cockroach GST causes IgE antibody responses and is associated with asthma. The data strongly support the view that the immune response to GST plays an important role in allergic diseases.