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Graphene and graphene oxide as a solid matrix for extraction of membrane and membrane-associated proteins
Microchimica Acta ( IF 5.7 ) Pub Date : 2018-01-19 , DOI: 10.1007/s00604-017-2658-5
Asad Uzzaman , Zhi Shang , Zhi Qiao , Cheng-Xi Cao , Hua Xiao

AbstractThe 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.

中文翻译:

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

摘要 由于固有的疏水性、动态差异和膜脂的抑制作用,膜蛋白的提取仍然是一个挑战。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。这里表明,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 分析进一步表明,与市售试剂盒相比,石墨烯(50-200 nm)和氧化石墨烯(50-200 nm)可以富集更多种类的膜蛋白。石墨烯进一步应用于富集正常细胞和癌细胞的膜蛋白,分别鉴定出1079和872种蛋白质,其中56.5%和60.5%为膜蛋白。特别是,241 种蛋白质在癌细胞中受到显着调节。15 种蛋白质的基因表达通过 qRT-PCR 验证,其中 4 种通过免疫分析进一步定量。这些数据共同表明,石墨烯具有改善膜蛋白提取的巨大潜力,因此可以服务于下游的癌症蛋白质组学。图形摘要以石墨烯和氧化石墨烯等二维碳纳米材料为固体基质,避免脂质双层干扰,提高膜和膜相关蛋白的提取效率。该策略将有利于下游膜蛋白质组学分析。这些数据共同表明,石墨烯具有改善膜蛋白提取的巨大潜力,因此可以服务于下游的癌症蛋白质组学。图形摘要以石墨烯和氧化石墨烯等二维碳纳米材料为固体基质,避免脂质双层干扰,提高膜和膜相关蛋白的提取效率。该策略将有利于下游膜蛋白质组学分析。这些数据共同表明,石墨烯在改善膜蛋白提取方面具有巨大潜力,因此可以服务于下游的癌症蛋白质组学。图形摘要以石墨烯和氧化石墨烯等二维碳纳米材料为固体基质,避免脂质双层干扰,提高膜和膜相关蛋白的提取效率。该策略将有利于下游膜蛋白质组学分析。
更新日期:2018-01-19
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