Graphene and graphene oxide as a solid matrix for extraction of membrane and membrane-associated proteins
Graphene and graphene oxide as a solid matrix for extraction of membrane and membrane-associated proteins
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石墨烯和氧化石墨烯作为固体基质用于提取膜和膜相关蛋白
DOI：
10.1007/s00604-017-2658-5
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发表时间：
2018
影响因子：
5.7
通讯作者：
Xiao Hua
中科院分区：
文献类型：
--
作者：
Uzzaman Asad;Shang Zhi;Qiao Zhi;Cao Cheng-Xi;Xiao Hua
由于膜蛋白的疏水性、动力学差异以及膜脂的抑制作用，膜蛋白的提取一直是一个难题。具有高表面积的纳米材料具有疏水-疏水脂质相互作用的能力。这里显示，溶解在增溶缓冲液中的石墨烯和氧化石墨烯都是有效提取膜蛋白的可行吸附剂。LC-MS/MS分析进一步显示，石墨烯（50-200 nm）和氧化石墨烯（50-200 nm）可以富集比市售试剂盒更多种类的膜蛋白。石墨烯进一步应用于正常细胞和癌细胞膜蛋白的富集，分别鉴定了1079和872个蛋白，其中56.5%和60.5%是膜蛋白。特别是，241种蛋白质在癌细胞中受到显著调节。通过qRT-PCR验证15种蛋白质的基因表达，并通过免疫测定进一步定量其中4种蛋白质。这些数据共同表明，石墨烯有很大的潜力，以改善膜蛋白的提取，从而可以为下游癌症蛋白质组学。图形摘要二维碳纳米材料，包括石墨烯和氧化石墨烯，被用作固体基质，以避免脂质双层干扰，提高膜和膜相关蛋白质的提取效率。该策略将有利于下游膜蛋白质组学分析。
The extraction of membrane proteins remain a challenge due to innate hydrophobicity, dynamic discrepancy, and restrain effect of membrane lipids. Nanomaterials with high surface area have competency of hydrophobic-hydrophobic lipid interactions. It is shown here that both graphene and graphene oxide dissolved in solubilization buffer are viable sorbents for efficient extraction of membrane proteins. LC-MS/MS analysis further revealed that graphene (50–200 nm) and graphene oxide (50–200 nm) can enrich more kinds of membrane proteins than a commercially available kit. Graphene was further applied to the enrichment of membrane proteins of normal cells as well as cancer cells, and 1079 and 872 proteins were identified, respectively, among which 56.5% and 60.5% were membrane proteins. In particular, 241 proteins were significantly regulated in cancer cells. Gene expression of 15 proteins was verified by qRT-PCR, and 4 of them were further quantified by immunoassay. These data collectively demonstrate that graphene has great potential to improve membrane protein extractions and thus can serve downstream cancer proteomics.Graphical abstractTwo dimensional carbon nanomaterials, including graphene and graphene oxide, were employed as solid matrix to avoid lipid bilayer interference and enhance the extraction efficiency of membrane and membrane associated proteins. The strategy will benefit downstream membrane proteomics analysis.