Selective enrichment and desalting of hydrophilic peptides using graphene oxide
Selective enrichment and desalting of hydrophilic peptides using graphene oxide
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使用氧化石墨烯选择性富集和脱盐亲水肽
DOI：
10.1016/j.jchromb.2016.05.044
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发表时间：
2016-08-01
影响因子：
3
通讯作者：
Chen, Ping
中科院分区：
文献类型：
--
作者：
Jiang, Miao;Qi, Linyu;Chen, Ping
组织中多肽的种类繁多，丰度低，给多肽的分离和鉴定带来了很大的困难，这不利于多肽组学的发展。RP-HPLC利用小分子的疏水性对其进行纯化，以其快速、重现性好、分辨率高的特点在多肽的分离和富集中得到了广泛的应用。然而，RP-HPLC需要仪器和昂贵的C18色谱柱，其样品容量也有限。最近，氧化石墨烯已被应用于氨基酸的吸附。然而，氧化石墨烯对多肽的富集效率和选择性尚不清楚。本研究比较了氧化石墨烯和RP-C18基质对胰蛋白酶化α -肌动蛋白以及垂体和海马组织提取物的吸附效率和选择性。对于α -肌动蛋白，总胰蛋白酶化产物和被氧化石墨烯和C18基质吸附的产物有相似的洗脱峰。而氧化石墨烯吸附的多肽亲水峰高于C18基质吸附的多肽。结果表明，氧化石墨烯富集的多肽大部分在低浓度的有机溶剂中被洗脱，而RP-C18基质吸附的多肽大部分在相对高浓度的有机溶剂中被洗脱。此外，质谱分析表明，垂体样品中氧化石墨烯富集了495个多肽，RP-C18基质富集了447个多肽，海马样品中分别富集了333个和243个多肽。卤汁值分析表明，与RP-C18基质相比，氧化石墨烯对高亲水性肽具有更强的吸附能力。此外，这两种方法的结合可以显著增加鉴定肽的数量和预测蛋白前体的数量。本研究为氧化石墨烯在组织多肽富集过程中的作用提供了新的思路，对多肽组学研究具有重要意义。(C) 2016 Elsevier B.V.版权所有
The wide variety and low abundance of peptides in tissue brought great difficulties to the separation and identification of peptides, which is not in favor of the development of peptidomics. RP-HPLC, which could purify small molecules based on their hydrophobicity, has been widely used in the separation and enrichment of peptide due to its fast, good reproducibility and high resolution. However, RP-HPLC requires the instrument and expensive C18 column and its sample capacity is also limited. Recently, graphene oxide has been applied to the adsorption of amino acids. However, the enrichment efficiency and selectivity of graphene oxide for peptides remain unclear. In this study, the adsorption efficiency and selectivity of graphene oxide and RP-C18 matrix were compared on trypsinized alpha-actin and also on tissue extracts from pituitary gland and hippocampus. For alpha-actin, there exhibit similar elution peaks for total trypsinized products and those adsorpted by GO and C18 matrix. But peptides adsorbed by GO showed the higher hydrophilic peaks than which adsorbed by C18 matrix. The resulted RP-HPLC profile showed that most of peptides enriched by graphene oxide were eluted at low concentration of organic solvent, while peptides adsorbed by RP-C18 matrix were mostly eluted at relatively high concentration. Moreover, mass spectrometry analysis suggested that, in pituitary sample, there were 495 peptides enriched by graphene oxide, 447 peptides enriched by RP-C18 matrix while in hippocampus sample 333 and 243 peptides respectively. The GRAVY value analysis suggested that the graphene oxide has a stronger adsorption for highly hydrophilic peptides compared to the RP-C18 matrix. Furthermore, the combination of these two methods could notably increase the number of identification peptides but also the number of predicted protein precursors. Our study provided a new thought to the role of graphene oxide during the enrichment of peptides from tissue which should be useful for peptidomics study. (C) 2016 Elsevier B.V. All rights reserved.