Design and synthesis of an immobilized metal affinity chromatography and metal oxide affinity chromatography hybrid material for improved phosphopeptide enrichment
Design and synthesis of an immobilized metal affinity chromatography and metal oxide affinity chromatography hybrid material for improved phosphopeptide enrichment
复制标题
设计和合成固定化金属亲和层析和金属氧化物亲和层析混合材料以改善磷酸肽富集
DOI：
10.1016/j.chroma.2017.05.025
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发表时间：
2017-07-07
影响因子：
4.1
通讯作者：
Yan, Fang
中科院分区：
文献类型：
--
作者：
Yang, Da-Song;Ding, Xi-Ying;Yan, Fang
蛋白质的可逆磷酸化是最重要的翻译后修饰之一，而磷酸肽的低丰度和非磷酸化肽的信号抑制给检测带来了困难。因此，从高度复杂的混合物中选择性地浓缩磷酸肽对于基于MS的磷酸蛋白质组分析是至关重要的。尽管已经开发了各种策略，但没有一种方法能够完全覆盖整个磷酸蛋白质组。金属氧化物亲和层析(MOAC)对单磷酸多肽具有较好的富集性，而固定化金属亲和层析(IMAC)对多个磷酸多肽的富集性较差。在本研究中，我们首次成功地合成了iMac和MOAc杂化材料Fe3O4@nSiO(2)@mSiO(2)/Ti02-Ti4+纳米颗粒，以富集单和多个磷酸化物种的优势。首先在介孔二氧化硅表面包覆二氧化钛，然后在其表面接枝甲基膦酸三羟基丙酯(THPMP)来螯合Ti4+离子。这种新型的杂化材料具有高比表面积(179.3 m(2)/g)，对酪蛋白的标准胰酶消化具有良好的吸附容量(133 mg/g)，基于该材料的方法也表现出良好的灵敏度(4pmoL)。将所合成的Fe3O4@nSiO(2)@mSiO(2)/Ti02-Ti4+微球用于从复杂的生物样品中选择性富集磷酸肽，成功地从脱脂牛奶中富集了7个单磷酸肽和8个多磷酸肽，明显优于单一IMAC或MOAC方法。这些结果表明，Fe3O4@nSiO(2)@mSiO(2)/Ti02-Ti4+微球在基于MS的磷酸蛋白质组学中具有潜在的应用前景，这一工作有望开辟一种将各种方法的优点结合在一种材料中有效丰富磷酸化多肽的策略。(C)2017爱思唯尔B.V.保留所有权利。
Reversible phosphorylation of proteins is one of the most important post-translational modifications, while the detection of phosphopeptides is difficult due to their low abundance and the signal suppression of nonphosphorylated peptides. Therefore, selective enrichment of phosphopeptides from highly complicated mixtures is vital for MS-based phosphoproteome analysis. Despite various strategies have been developed, there is no single method that is capable of providing full coverage of the whole phosphoproteome. Metal oxide affinity chromatography (MOAC) enrichment preferably singly phosphopeptides, whereas immobilized metal affinity chromatography (IMAC) enrichment bias towards multiply phosphopeptides. In this study, first example of IMAC and MOAC hybrid material, Fe3O4@nSiO(2)@mSiO(2)/TiO2-Ti4+ nanoparticles were successfully synthesized for the enrichment of phosphopeptides with the aim to combining their advantages for enrich both mono-and multi-phosphorylated species. The TiO2 was firstly coated on the surface of mesoporous silica and then grafted with 3-(trihydroxysilyl)propyl methylphosphonate (THPMP) to chelate Ti4+ ions. This novel type of hybird material with high surface areas (179.3 m(2)/g) exhibited good adsorption capacity (133 mg/g) towards standard tryptic digest of,casein and the method based on this material also showed good sensitivity (4 pmol). The synthesized Fe3O4@nSiO(2)@mSiO(2)/TiO2-Ti4+ microspheres were further used to selectively enrich phosphopeptides from complex biosamples, seven mono-phosphopeptides and eight multi-phosphopeptides were successfully enriched from nonfat milk which is much better than single IMAC or MOAC strategy. Those results indicated that Fe3O4@nSiO(2)@mSiO(2)/TiO2-Ti4+ microspheres have potential applications in MS based phosphoproteomics to enlarge phosphoproteomics coverage and this work was expected to open up a promising strategy which combined the advantages of various methods in one material for effective enrich phosphorylated peptides. (C) 2017 Elsevier B.V. All rights reserved.