Detecting Proteins Glycosylation by a Homogeneous Reaction System with Zwitterionic Gold Nanoclusters
Detecting Proteins Glycosylation by a Homogeneous Reaction System with Zwitterionic Gold Nanoclusters
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通过两性离子金纳米簇均相反应系统检测蛋白质糖基化
DOI：
10.1021/acs.analchem.6b04880
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发表时间：
2017
影响因子：
7.4
通讯作者：
Wang Fangjun
中科院分区：
文献类型：
--
作者：
Li Jinan;Liu Jing;Liu Zheyi;Tan Yuan;Liu Xiaoyan;Wang Fangjun
均匀的金纳米团簇（Au NCs）由于其独特的物理和化学性质，在药物输送、化学传感、生物检测和生物标记等方面得到了广泛的应用。然而，很少有人注意到它们在检测蛋白质翻译后修饰中的应用。在此，我们描述了一个均相反应系统的发展与水溶性两性离子Au NC捕获糖肽从复杂的生物样品。Au NCs的独特特性，如它们的分子样性质、在水溶液中优异的均匀性、有机溶剂响应性沉淀以及仅4.5 h的容易制备，有助于高效率和高通量地捕获靶向糖肽。与传统的非均相固相吸附系统相比，糖基化位点的数量提高了35%。最后，标准糖蛋白（IgG）的MS检测限低至50阿莫尔，仅从60 μg小鼠肝蛋白中鉴定出713种糖蛋白的1576个糖基化位点。数据可通过ProteomeXchange获得，标识符为PXD 005635。
Homogeneous gold nanoclusters (Au NCs) have been widely utilized in drug delivery, chemical sensing, bioassays, and biolabeling due to their unique physical and chemical properties. However, little attention has been paid to their application in detecting protein post-translational modifications. Herein, we describe the development of a homogeneous reaction system with water-soluble zwitterionic Au NCs to capture glycopeptides from complex biological samples. The unique characteristics of Au NCs, such as their molecular-like properties, the excellent homogeneity in aqueous solution, the organic solvent responsive precipitation, and the easy preparation in only 4.5 h, contribute to the high efficiency and high throughput for capturing the targeted glycopeptides. Compared with the conventional heterogeneous system with solid-state adsorbents, the number of characterized glycosylation sites was improved by 35%. Finally, an MS detection limit as low as 50 amol was achieved for the standard glycoprotein (IgG), and 1576 glycosylation sites from 713 glycoproteins were identified from only 60 μg of mouse liver protein. Data are available via ProteomeXchange with identifier PXD005635.