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Phosphonate-modified metal oxides for the highly selective enrichment of phosphopeptides

机译：膦酸酯改性金属氧化物，用于磷酸肽的高选择性富集

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摘要

Metal oxide affinity chromatography has become one of the most widely used strategies for phosphopeptide enrichment prior to mass spectrometry analysis, but some defects still exist in this approach due to the complex physical and chemical properties of the metal oxide surface. Although the simultaneous phosphorylation of adjacent amino acids may greatly affect the bioactivity of the protein, there are few reports on the specific enrichment of multi-phosphopeptides. In this work, we report a highly selective enrichment method for capturing phosphopeptides or multi-phosphopeptides based on phosphonate-modified metal oxides. Compounds with different numbers of phosphate groups were adsorbed on the surface of ZrO2 and TiO2 to obtain phosphonate-modified metal oxides. Among them, phosphoric acid modified metal oxides (1P-ZrO2 and 1P-TiO2) could significantly enhance their selectivity towards phosphopeptides; and alendronate-modified metal oxides (2P-ZrO2 and 2P-TiO2) showed high selectivity for the enrichment of multi-phosphopeptides. In addition, the detection sensitivity was greatly improved by using these novel materials. The mechanism of the specific enrichment was considered to be ligand exchange and blocking of strong adsorption sites by the compounds containing the phosphate group. Finally, tryptic digests of proteins of human Jurkat-T cell lysate were further used to demonstrate the selectivity and specificity of ZrO2, 1P-ZrO2 and 2P-ZrO2.
机译：金属氧化物亲和色谱法已成为质谱分析之前最广泛使用的磷酸富集的策略之一，但由于金属氧化物表面的复杂物理和化学性质，这种方法仍然存在一些缺陷。虽然相邻氨基酸的同时磷酸化可能极大地影响蛋白质的生物活性，但仍有关于多磷酸肽的比富集的报道。在这项工作中，我们报告了一种高度选择性的富集方法，用于基于膦酸盐改性的金属氧化物捕获磷酸肽或多磷酸酯。具有不同数量的磷酸盐基团的化合物被吸附在ZrO2和TiO 2的表面上，得到膦酸盐改性的金属氧化物。其中，磷酸改性金属氧化物（1p-ZrO2和1P-TiO 2）可以显着提高它们对磷酸肽的选择性;和醛膦改性的金属氧化物（2P-ZrO2和2P-TiO 2）显示出高选择性的多磷酸肽的富集。此外，通过使用这些新颖的材料大大提高了检测灵敏度。具体富集的机制被认为是配体交换和含有磷酸盐基团的化合物的强吸附位点。最后，进一步用于证明ZrO2，1p-ZrO2和2P-ZrO2的选择性和特异性的人为Jurkat-T细胞裂解物的胰蛋白质毒素消化。

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  • 来源
    《RSC Advances》 |2015年第11期|共10页
  • 作者

    Li Xiao-Shui; Chen Xi; Yuan Bi-Feng; Feng Yu-Qi;

  • 作者单位

    Wuhan Univ Dept Chem Key Lab Analyt Chem Biol &

    Med Minist Educ Wuhan 430072 Peoples R China;

    Wuhan Inst Biotechnol Wuhan 430072 Peoples R China;

    Wuhan Univ Dept Chem Key Lab Analyt Chem Biol &

    Med Minist Educ Wuhan 430072 Peoples R China;

    Wuhan Univ Dept Chem Key Lab Analyt Chem Biol &

    Med Minist Educ Wuhan 430072 Peoples R China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 化学;
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