Exploring Proteins in Anopheles gambiae Male and Female Antennae through MALDI Mass Spectrometry Profiling
Exploring Proteins in Anopheles gambiae Male and Female Antennae through MALDI Mass Spectrometry Profiling
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DOI：
10.1371/journal.pone.0002822
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发表时间：
2008-07-30
期刊：
影响因子：
3.7
通讯作者：
Turillazzi, Stefano
中科院分区：
文献类型：
--
作者：
Dani, Francesca R.;Francese, Simona;Turillazzi, Stefano
MALDI谱和成像质谱（IMS）是直接分析生物组织中肽和小分子蛋白质的新技术。在这项工作中，我们将其应用到冈比亚按蚊触角的研究，目的是分析参与嗅觉周围事件的可溶性蛋白质的表达。通过对单个触角的直接分析和提取物的分析获得的MALDI光谱显示出相似的轮廓，尽管通过分析获得的光谱具有更丰富的离子群和更高的信噪比。雄性和雌性触角显示出不同的蛋白质谱。还进行了MALDI成像实验，并观察到一些蛋白质的定位差异。通过高分辨率测量和自顶向下的MS/MS实验鉴定了两种蛋白质。一个8 kDa的蛋白质只存在于雄性触角匹配的一个未注释的序列的An。通过双向电泳、质谱和质谱/质谱分析，证实了冈比亚红球藻基因组中存在气味结合蛋白9（OBP-9）。这项工作表明，MALDI MS分析是一种适合于分析昆虫附录中的小分子量和中等分子量蛋白质的技术，并允许获得几个标本的数据，可以调查组间的差异。感兴趣的蛋白质可以通过其他互补的MS方法来鉴定。
MALDI profiling and imaging mass spectrometry (IMS) are novel techniques for direct analysis of peptides and small proteins in biological tissues. In this work we applied them to the study of Anopheles gambiae antennae, with the aim of analysing expression of soluble proteins involved in olfaction perireceptor events. MALDI spectra obtained by direct profiling on single antennae and by the analysis of extracts, showed similar profiles, although spectra obtained through profiling had a richer ion population and higher signal to noise ratio. Male and female antennae showed distinct protein profiles. MALDI imaging experiments were also performed and differences were observed in the localization of some proteins. Two proteins were identified through high resolution measurement and top-down MS/MS experiments. A 8 kDa protein only present in the male antennae matched with an unannotated sequence of the An. gambiae genome, while the presence of odorant binding protein 9 (OBP-9) was confirmed through experiments of 2-DE, followed by MS and MS/MS analysis of digested spots. This work shows that MALDI MS profiling is a technique suitable for the analysis of proteins of small and medium MW in insect appendices, and allows obtaining data for several specimens which can be investigated for differences between groups. Proteins of interest can be identified through other complementary MS approaches.