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Preparation of graphene-hafnium oxide composite for selective enrichment and analysis of phosphopeptides

机译：石墨烯 - 氧化铪复合材料的选择性富集和分析磷酸肽分析

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摘要

The reversible phosphorylation of proteins plays a crucial role in many regulatory processes. During the last decade, there has been considerable interest in the development of new methods for the identification of phosphorylation sites to allow for the comprehensive analysis of protein phosphorylation processes. However, the development of sensitive methods for the detection of trace quantities of phosphopeptides remains a significant challenge. In this study, we have prepared a novel graphene- hafnium oxide composite (GHOC) capable of enriching phosphopeptides and its application for the enrichment of phosphopeptides prior to their analysis by matrix-assisted laser desorption/ionization mass spectrometry (MALDI - TOF/MS). The GHOC was prepared in a facile step using a hydrothermal reaction. The surface morphology of the resultant materials was analyzed in its compound-bound form by TEM, SAED and XRD. According to theses results, the GHOC had an increased surface area resulting from the template of graphene and the modification of hafnium oxide which possess high specificity toward phosphopeptides. Several complex samples (e.g. α-and b-casein, mixtures of b-casein and bovine serum albumin, and nonfat milk tryptic digest) were used to test the enrichment capability of the GHOC, and the results demonstrated that this material exhibited better selectivity towards mono-and multi-phosphorylated peptides compared with the graphene-TiO2 composite (GTOC) and graphene- ZrO2 composite (GZOC). Furthermore, MALDI-TOF/MS experiments revealed no interference from nonphosphopeptides. The results demonstrate that our newly developed GHOC shows high specificity for the enrichment of phosphopeptides from biological samples and could therefore be applied in the field of phosphoproteomics.
机译：蛋白质的可逆磷酸化在许多监管过程中起着至关重要的作用。在过去十年中，对鉴定磷酸化位点的新方法进行了相当兴趣，以允许综合分析蛋白质磷酸化过程。然而，用于检测痕量磷酸肽的敏感方法的发展仍然是一个重大挑战。在这项研究中，我们制备了一种新的石墨烯 - 氧化铪复合物（GHOC），其能够在通过基质辅助激光解吸/电离质谱（MALDI - TOF / MS）分析之前富集磷酸肽及其富集磷酸肽的富集。使用水热反应在容纳步骤中制备GHOC。通过TEM，SAED和XRD分析所得材料的表面形态。根据结果​​，GHOC具有由石墨烯模板和铪氧化铪的改性产生的表面积增加，该氧化铪对磷酸肽具有高特异性。使用几种复杂样品（例如α-和B-酪蛋白，B-酪蛋白和牛血清白蛋白的混合物，并且非比牛奶胰蛋白酶消化）测试GHOC的富集能力，结果表明该材料表现出更好的选择性与石墨烯-TiO2复合物（GTOC）和石墨烯-ZRO2复合物（GZOC）相比单磷酸化肽。此外，MALDI-TOF / MS实验揭示了非磷代肽的干扰。结果表明，我们的新开发的GHOC表明了对生物样品的富集磷酸肽的富集具有高特异性，因此可以应用于磷蛋白蛋白质的领域。

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  • 来源
    《RSC Advances》 |2015年第109期|共8页
  • 作者

    Xuan Huang; Junping Wang; Junying Wang;

  • 作者单位

    Key Laboratory of Food Nutrition and Safety Ministry of Education Tianjin Key Laboratory of Food Nutrition and Safety Tianjin University of Science and Technology Tianjin 300457 China.;

    Key Laboratory of Food Nutrition and Safety Ministry of Education Tianjin Key Laboratory of Food Nutrition and Safety Tianjin University of Science and Technology Tianjin 300457 China.;

    Biotechnology Research Institute Chinese Academy of Agricultural Sciences Beijing 10081 China.;

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  • 正文语种 eng
  • 中图分类 化学;
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