A Comparative Study of Electrostatic Repulsion-Hydrophilic Interaction Chromatography (ERLIC) versus SCX-IMAC-Based Methods for Phosphopeptide Isolation/Enrichment

被引：80

作者：

Gan, Chee Sian ^{[1
]}

Guo, Tiannan ^{[1
,2
]}

Zhang, Huoming ^{[1
]}

Lim, Sai Kiang ^{[3
]}

Sze, Siu Kwan ^{[1
]}

机构：

[1] Nanyang Technol Univ, Sch Biol Sci, Singapore 637551, Singapore

[2] Natl Canc Ctr Singapore, Div Med Sci, Singapore 169610, Singapore

[3] Inst Med Biol, Singapore 138648, Singapore

来源：

JOURNAL OF PROTEOME RESEARCH | 2008年 / 7卷 / 11期

关键词：

Phosphopeptide enrichment; EGFR signaling; A431 carcinoma cells; WAX; IMAC; ERLIC; SCX;

D O I：

10.1021/pr800473j

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Electrostatic repulsion-hydrophilic interaction chromatography (ERLIC) has been introduced recently for phosphopeptide enrichment. Here we compared ERLIC with the well-established SCX-IMAC for identifying phosphopeptides in EGF-treated A431 cells. The ERLIC approach detected a higher number of phosphopeptides (17 311) than SCX-IMAC (4850), but it only detected 926 unique phosphopeptides compared to 1315 in SCX-IMAC. Only 12% unique phosphopeptides were common to both approaches, suggesting that more comprehensive phosphoproteomes could be generated by complementing SCX-IMAC with ERLIC.

引用

页码：4869 / 4877

页数：9

共 34 条

[1]

ALPERT A, 2008, ABRF C

[2] Electrostatic repulsion hydrophilic interaction chromatography for isocratic separation of charged solutes and selective isolation of phosphopeptides [J].

Alpert, Andrew J. .

ANALYTICAL CHEMISTRY, 2008, 80 (01) :62-76

[3] A curated compendium of phosphorylation motifs [J].

Amanchy, Ramars ;

Periaswamy, Balamurugan ;

Mathivanan, Suresh ;

Reddy, Raghunath ;

Tattikota, Sudhir Gopal ;

Pandey, Akhilesh .

NATURE BIOTECHNOLOGY, 2007, 25 (03) :285-286

[4] Large-scale characterization of HeLa cell nuclear phosphoproteins [J].

Beausoleil, SA ;

Jedrychowski, M ;

Schwartz, D ;

Elias, JE ;

Villén, J ;

Li, JX ;

Cohn, MA ;

Cantley, LC ;

Gygi, SP .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (33) :12130-12135

[5] A probability-based approach for high-throughput protein phosphorylation analysis and site localization [J].

Beausoleil, Sean A. ;

Villen, Judit ;

Gerber, Scott A. ;

Rush, John ;

Gygi, Steven P. .

NATURE BIOTECHNOLOGY, 2006, 24 (10) :1285-1292

[6] Reproducible isolation of distinct, overlapping segments of the phosphoproteome [J].

Bodenmiller, Bernd ;

Mueller, Lukas N. ;

Mueller, Markus ;

Domon, Bruno ;

Aebersold, Ruedi .

NATURE METHODS, 2007, 4 (03) :231-237

[7] Target-decoy search strategy for increased confidence in large-scale protein identifications by mass spectrometry [J].

Elias, Joshua E. ;

Gygi, Steven P. .

NATURE METHODS, 2007, 4 (03) :207-214

[8] Phosphoproteome analysis by mass spectrometry and its application to Saccharomyces cerevisiae [J].

Ficarro, SB ;

McCleland, ML ;

Stukenberg, PT ;

Burke, DJ ;

Ross, MM ;

Shabanowitz, J ;

Hunt, DF ;

White, FM .

NATURE BIOTECHNOLOGY, 2002, 20 (03) :301-305

[9] PHOSIDA (phosphorylation site database): management, structural and evolutionary investigation, and prediction of phosphosites [J].

Gnad, Florian ;

Ren, Shubin ;

Cox, Juergen ;

Olsen, Jesper V. ;

Macek, Boris ;

Oroshi, Mario ;

Mann, Matthias .

GENOME BIOLOGY, 2007, 8 (11)

[10] Quantitative phosphoproteomics applied to the yeast pheromone signaling pathway [J].

Gruhler, A ;

Olsen, JV ;

Mohammed, S ;

Mortensen, P ;

Færgeman, NJ ;

Mann, M ;

Jensen, ON .

MOLECULAR & CELLULAR PROTEOMICS, 2005, 4 (03) :310-327

← 1 2 3 4 →