Effective Enrichment and Mass Spectrometry Analysis of Phosphopeptides Using Mesoporous Metal Oxide Nanomaterials

被引：71

作者：

Nelson, Cory A. ^{[1
,2
]}

Szczech, Jeannine R. ^{[1
]}

Dooley, Chad J. ^{[1
]}

Xu, Qingge ^{[2
]}

Lawrence, Matthew J. ^{[2
]}

Zhu, Haoyue ^{[1
]}

Jin, Song ^{[1
]}

Ge, Ying ^{[2
]}

机构：

[1] Univ Wisconsin, Dept Chem, Madison, WI 53706 USA

[2] Univ Wisconsin, Sch Med & Publ Hlth, Human Prote Program, Dept Physiol, Madison, WI 53706 USA

来源：

ANALYTICAL CHEMISTRY | 2010年 / 82卷 / 17期

基金：

美国国家卫生研究院;

关键词：

ELECTRON-CAPTURE DISSOCIATION; HIGHLY SELECTIVE ENRICHMENT; TITANIA THIN-FILMS; PHOSPHORYLATED PEPTIDES; PROTEIN-PHOSPHORYLATION; PHOSPHOPROTEOME ANALYSIS; AFFINITY-CHROMATOGRAPHY; AQUEOUS-SOLUTIONS; FEMTOMOLE LEVEL; ZIRCONIUM-OXIDE;

D O I：

10.1021/ac100877a

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Mass spectrometry (MS)-based phosphoproteomics remains challenging due to the low abundance of phosphoproteins and substoichiometric phosphotylation. This demands better methods to effectively enrich phosphoproteins/peptides prior to MS analysis. We have previously communicated the first use of mesoporous zirconium dioxide (ZrO2) nanomaterials for effective phosphopeptide enrichment. Here, we present the full report including the synthesis, characterization, and application of mesoporous titanium dioxide (TiO2), ZrO2, and hafnium dioxide (HfO2) in phosphopeptide enrichment and MS analysis. Mesoporous ZrO2 and HfO2 are demonstrated to be superior to TiO2 for phosphopeptide enrichment from a complex mixture with high specificity (>99%), which could almost be considered as a "purification", mainly because of the extremely large active surface area of mesoporous nanomaterials. A single enrichment and Fourier transform MS analysis of phosphopeptides digested from a complex mixture containing 7% of alpha-casein identified 21 out of 22 phosphorylation sites for alpha-casein. Moreover, the mesoporous ZrO2 and HfO2 can be reused after a simple solution regeneration procedure with comparable enrichment performance to that of fresh materials. Mesoporous ZrO2 and HfO2 nanomaterials hold great promise for applications in MS-based phosphoproteomics.

引用

页码：7193 / 7201

页数：9

共 61 条

[1] In Vivo Phosphorylation Site Mapping in Mouse Cardiac Troponin I by High Resolution Top-Down Electron Capture Dissociation Mass Spectrometry: Ser22/23 Are the Only Sites Basally Phosphorylated [J].