DICP OpenIR
Robust phosphoproteome enrichment using monodisperse microsphere-based immobilized titanium (IV) ion affinity chromatography
Zhou, Houjiang1,2,3; Ye, Mingliang2; Dong, Jing2; Corradini, Eleonora1,3; Cristobal, Alba1,3; Heck, Albert J. R.1,3; Zou, Hanfa2; Mohammed, Shabaz1,3
刊名NATURE PROTOCOLS
2013-03-01
DOI10.1038/nprot.2013.010
8期:3页:461-480
收录类别SCI
文章类型Article
WOS标题词Science & Technology ; Life Sciences & Biomedicine
类目[WOS]Biochemical Research Methods
研究领域[WOS]Biochemistry & Molecular Biology
关键词[WOS]MASS-SPECTROMETRIC ANALYSIS ; COMPLEX PROTEIN MIXTURES ; PHOSPHOPEPTIDE ENRICHMENT ; SACCHAROMYCES-CEREVISIAE ; PHOSPHORYLATED PEPTIDES ; MS ANALYSIS ; SITE LOCALIZATION ; POLYMER PARTICLES ; PROTEOMICS ; TANDEM
英文摘要Mass spectrometry (MS)-based proteomics has become the preferred tool for the analysis of protein phosphorylation. To be successful at such an endeavor, there is a requirement for an efficient enrichment of phosphopeptides. This is necessary because of the substoichiometric nature of phosphorylation at a given site and the complexity of the cell. Recently, new alternative materials have emerged that allow excellent and robust enrichment of phosphopeptides. These monodisperse microsphere-based immobilized metal ion affinity chromatography (IMAC) resins incorporate a flexible linker terminated with phosphonate groups that chelate either zirconium or titanium ions. The chelated zirconium or titanium ions bind specifically to phosphopeptides, with an affinity that is similar to that of other widely used metal oxide affinity chromatography materials (typically TiO2). Here we present a detailed protocol for the preparation of monodisperse microsphere-based Ti4+-IMAC adsorbents and the subsequent enrichment process. Furthermore, we discuss general pitfalls and crucial steps in the preparation of phosphoproteomics samples before enrichment and, just as importantly, in the subsequent mass spectrometric analysis. Key points such as lysis, preparation of the chromatographic system for analysis and the most appropriate methods for sequencing phosphopeptides are discussed. Bioinformatics analysis specifically relating to site localization is also addressed. Finally, we demonstrate how the protocols provided are appropriate for both single-protein analysis and the screening of entire phosphoproteomes. It takes similar to 2 weeks to complete the protocol: 1 week to prepare the Ti4+-IMAC material, 2 d for sample preparation, 3 d for MS analysis of the enriched sample and 2 d for data analysis.
语种英语
WOS记录号WOS:000317110600003
引用统计
被引频次:162[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/137632
专题中国科学院大连化学物理研究所
作者单位1.Univ Utrecht, Utrecht Inst Pharmaceut Sci, Bijvoet Ctr Biomol Res, Utrecht, Netherlands
2.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian, Peoples R China
3.Netherlands Prote Ctr, Utrecht, Netherlands
推荐引用方式
GB/T 7714
Zhou, Houjiang,Ye, Mingliang,Dong, Jing,et al. Robust phosphoproteome enrichment using monodisperse microsphere-based immobilized titanium (IV) ion affinity chromatography[J]. NATURE PROTOCOLS,2013,8(3):461-480.
APA Zhou, Houjiang.,Ye, Mingliang.,Dong, Jing.,Corradini, Eleonora.,Cristobal, Alba.,...&Mohammed, Shabaz.(2013).Robust phosphoproteome enrichment using monodisperse microsphere-based immobilized titanium (IV) ion affinity chromatography.NATURE PROTOCOLS,8(3),461-480.
MLA Zhou, Houjiang,et al."Robust phosphoproteome enrichment using monodisperse microsphere-based immobilized titanium (IV) ion affinity chromatography".NATURE PROTOCOLS 8.3(2013):461-480.
条目包含的文件
条目无相关文件。
个性服务
推荐该条目
保存到收藏夹
查看访问统计
导出为Endnote文件
谷歌学术
谷歌学术中相似的文章
[Zhou, Houjiang]的文章
[Ye, Mingliang]的文章
[Dong, Jing]的文章
百度学术
百度学术中相似的文章
[Zhou, Houjiang]的文章
[Ye, Mingliang]的文章
[Dong, Jing]的文章
必应学术
必应学术中相似的文章
[Zhou, Houjiang]的文章
[Ye, Mingliang]的文章
[Dong, Jing]的文章
相关权益政策
暂无数据
收藏/分享
所有评论 (0)
暂无评论
 

除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。