A robust new strategy for high-molecular-weight proteome research: a 2-hydroxyethyl agarose/polyacrylamide gel enhanced separation and ZnO-PMMA nanobeads assisted identification.
A robust new strategy for high-molecular-weight proteome research: a 2-hydroxyethyl agarose/polyacrylamide gel enhanced separation and ZnO-PMMA nanobeads assisted identification.
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高分子量蛋白质组研究的稳健新策略：2-羟乙基琼脂糖/聚丙烯酰胺凝胶增强分离和 ZnO-PMMA 纳米珠辅助鉴定。
DOI：
10.1016/j.talanta.2010.06.034
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发表时间：
2010
期刊：
影响因子：
6.1
通讯作者：
Pengyuan Yang
中科院分区：
文献类型：
--
作者：
Wenwen Shen;Chengpin Shen;H. Xiong;Haojie Lu;Pengyuan Yang
建立了一种新的基于质谱的高分子量蛋白质组研究方法。首先，首次设计了2-羟乙基琼脂糖/聚丙烯酰胺（HEAG/PAM）电泳凝胶，实现了一种操作简单的高分子量蛋白质的分离方法，具有高空间分辨率，良好的重现性和质谱兼容的银染。其次，氧化锌-聚甲基丙烯酸甲酯（ZnO-PMMA）纳米珠在这里被应用于富集和纯化的肽从HMW蛋白质。第三，通过基质辅助激光解吸/电离质谱（MALDI-MS）分析的肽与ZnO-PMMA纳米珠的存在下，和它们的MS信号显着增强。由于ZnO-PMMA纳米珠的高富集效率和耐盐能力以及信号增强能力，高分子量蛋白的鉴定成功率显著提高。我们相信，这种分析策略将启发和促进高分子量蛋白质组研究。
A new mass spectrometry based analysis strategy has been established here for high-molecular-weight (HMW) proteome research. First, a 2-hydroxyethyl agarose/polyacrylamide (HEAG/PAM) electrophoresis gel was designed for the first time to realize an easy-handling separation method with high spatial resolution for HMW proteins, good reproducibility and mass spectrometry-compatible sliver staining. Second, ZnO–polymethyl methacrylate (ZnO–PMMA) nanobeads were applied here for enriching and desalting the peptides from the HMW proteins. Third, the peptides were analyzed by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) with the presence of the ZnO–PMMA nanobeads, and their MS signals were enhanced markedly. The success rate of identification for HMW proteins was significantly increased due to high enriching efficiency and salt tolerance capability as well as signal enhancing capability of the ZnO–PMMA nanobeads. We believe that this analysis strategy will inspire and accelerate the HMW proteome studies.