Selective zirconium dioxide-based enrichment of phosphorylated peptides for mass spectrometric analysis
Selective zirconium dioxide-based enrichment of phosphorylated peptides for mass spectrometric analysis
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DOI：
10.1021/ac0522355
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发表时间：
2006-03-15
影响因子：
7.4
通讯作者：
Håkansson, K
中科院分区：
文献类型：
--
作者：
Kweon, HK;Håkansson, K
由于蛋白质磷酸化的动态性质和低化学计量，在通过质谱法表征之前，通常需要从蛋白水解混合物中富集磷酸化肽。在文献中已经提出了几种磷酸肽分离策略，包括固定化金属离子亲和色谱。然而，该技术的选择性和再现性差。近年来，二氧化钛基柱已被一些研究小组成功地用于磷酸肽富集。在这里，我们提出，据我们所知，第一次演示的实用程序的二氧化锆microtips磷酸肽分离质谱分析之前。这些微尖显示出与TiO 2微尖相似的整体性能。然而，更有选择性的分离单一磷酸化的肽，观察与氧化锆相比，TiO 2，而TiO 2优先富集多磷酸化的肽。因此，这两种色谱材料具有互补性质。对于α-和β-酪蛋白，Glu-C消化相比，胰蛋白酶消化时，结合TiO 2或ZrO 2磷酸肽富集没有明显的优势。
Due to the dynamic nature and low stoichiometry of protein phosphorylation, enrichment of phosphorylated peptides from proteolytic mixtures is often necessary prior to their characterization by mass spectrometry. Several phosphopeptide isolation strategies have been presented in the literature, including immobilized metal ion affinity chromatography. However, that technique suffers from poor selectivity and reproducibility. Recently, titanium dioxide-based columns have been successfully employed for phosphopeptide enrichment by several research groups. Here, we present, to our knowledge, the first demonstration of the utility of zirconium dioxide microtips for phosphopeptide isolation prior to mass spectrometric analysis. These microtips display similar overall performance as TiO2 microtips. However, more selective isolation of singly phosphorylated peptides was observed with ZrO2 compared to TiO2 whereas TiO2 preferentially enriched multiply phosphorylated peptides. Thus, these two chromatographic materials possess complementary properties. For alpha- and beta-casein, Glu-C digestion provided no evident advantage compared to trypsin digestion when combined with TiO2 or ZrO2 phosphopeptide enrichment.