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从纤维素分解真菌卷枝毛霉中鉴定一种壳聚糖酶的特性。

Characterization of an antigenic chitosanase from the cellulolytic fungus Chaetomium globosum.

机构信息

Ottawa-Carleton Institute of Chemistry, Carleton University, Ottawa, ON, Canada.

出版信息

Med Mycol. 2013 Apr;51(3):290-9. doi: 10.3109/13693786.2012.715246. Epub 2012 Sep 18.

Abstract

We are interested in identifying human fungal allergens and antigens from species common on water-damaged or damp building materials for use as marker proteins and diagnostic tests. The cellulolytic fungus Chaetomium globosum is common on damp materials in the building environment worldwide. ELISA and immunoblotting tests identified two related proteins of molecular weights 45 and 47 kDa which were identified as fungal antigens found on spore surfaces and in culture filtrate. The sequences were determined by liquid chromatography tandem mass spectrometry (LC-MS/MS), which indicated that the two proteins were chitosanases, confirmed by enzyme assay. The 47 kDa protein was not glycosylated and had an acidic pI of 4.5. These proteins have not been reported from other fungi and similar antigens were not seen in other fungi common in buildings. The production of polyclonal antibodies in rabbits showed the antigenicity of the target proteins and confirmed they were not artifacts of the isolation process. The proteins isolated are useful biomarkers for the detection of C. globosum in the building environment.

摘要

我们有兴趣从常见于水损坏或潮湿建筑材料上的物种中鉴定人类真菌过敏原和抗原,以用作标记蛋白和诊断测试。纤维素分解真菌Chaetomium globosum 在世界各地的建筑环境中常见于潮湿材料上。ELISA 和免疫印迹测试鉴定出两种相关的分子量为 45 和 47 kDa 的蛋白质,它们被鉴定为存在于孢子表面和培养滤液中的真菌抗原。通过液相色谱串联质谱(LC-MS/MS)确定了序列,表明这两种蛋白质是壳聚糖酶,通过酶测定得到证实。47 kDa 蛋白未糖基化,等电点为 4.5。这些蛋白质尚未从其他真菌中报道,在建筑物中常见的其他真菌中也未观察到类似的抗原。用兔子产生的多克隆抗体显示了目标蛋白的抗原性,并证实它们不是分离过程的人工制品。分离出的蛋白质是检测建筑环境中 C. globosum 的有用生物标志物。

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