Phosphorylation Analysis of Primary Human T Lymphocytes Using Sequential IMAC and Titanium Oxide Enrichment

被引：65

作者：

Carrascal, Montserrat ^{[1
]}

Ovefletro, David ^{[1
]}

Casas, Vanessa ^{[1
]}

Gay, Marina ^{[1
]}

Ablan, Joaquin ^{[1
]}

机构：

[1] Fac Med, CSIC UAB Prote Lab, IIBB, IDIBAPS, Bellaterra 08193, Spain

来源：

JOURNAL OF PROTEOME RESEARCH | 2008年 / 7卷 / 12期

关键词：

Phosphoproteomics; T lymphocytes; immune response; IMAC; titanium oxide; LC-MSn; LymPHOS database;

D O I：

10.1021/pr800500r

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

T lymphocytes mediate cellular and humoral defense against foreign bodies or autoantigens. An understanding of T-cell information processing furthers studies of the immunological response. We describe a large-scale phosphorylation analysis of primary T cells using a multidimensional separation strategy, involving preparative SDS-PAGE for prefractionation, in-gel digestion and sequential phosphopeptide enrichment using IMAC and TiO2. A total of 281 phosphorylation sites (197 of high confidence, Ascore > 15), mapping to 204 human gene sequences, were identified by LC-MSn analysis in an LTQ linear ion trap. Subsequently, we created the LymPHOS database (http://lymphos.org), which links mass spectrometric peptide information to phosphorylation sites and phosphoprotein sequences.

引用

页码：5167 / 5176

页数：10

共 59 条

[1] 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

[2] ISOLATION OF PHOSPHOPROTEINS BY IMMOBILIZED METAL (FE-3+) AFFINITY-CHROMATOGRAPHY [J].

ANDERSSON, L ;

PORATH, J .

ANALYTICAL BIOCHEMISTRY, 1986, 154 (01) :250-254

[3] The effects of mass accuracy, data acquisition speed, and search algorithm choice on peptide identification rates in phosphoproteomics [J].

Bakalarski, Corey E. ;

Haas, Wilhelm ;

Dephoure, Noah E. ;

Gygi, Steven P. .

ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2007, 389 (05) :1409-1419

[4] 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

[5] PhosphoPep-a phosphoproteome resource for systems biology research in Drosophila Kc167 cells [J].

Bodenmiller, Bernd ;

Malmstrom, Johan ;

Gerrits, Bertran ;

Campbell, David ;

Lam, Henry ;

Schmidt, Alexander ;

Rinner, Oliver ;

Mueller, Lukas N. ;

Shannon, Paul T. ;

Pedrioli, Patrick G. ;

Panse, Christian ;

Lee, Hoo-Keun ;

Schlapbach, Ralph ;

Aebersold, Ruedi .

MOLECULAR SYSTEMS BIOLOGY, 2007, 3 (1)

[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] Robust phosphoproteomic profiling of tyrosine phosphorylation sites from human T cells using immobilized metal affinity chromatography and tandem mass spectrometry [J].

Brill, LM ;

Salomon, AR ;

Ficarro, SB ;

Mukherji, M ;

Stettler-Gill, M ;

Peters, EC .

ANALYTICAL CHEMISTRY, 2004, 76 (10) :2763-2772

[8] Combining protein-based IMAC, peptide-based IMAC, and MudPIT for efficient phosphoproteomic analysis [J].

Cantin, Greg T. ;

Yi, Wei ;

Lu, Bingwen ;

Park, Sung Kyu ;

Xu, Tao ;

Lee, Jiing-Dwan ;

Yates, John R., III .

JOURNAL OF PROTEOME RESEARCH, 2008, 7 (03) :1346-1351

[9] Quantitative phosphoproteomic analysis of the tumor necrosis factor pathway [J].

Cantin, GT ;

Venable, JD ;

Cociorva, D ;

Yates, JR .

JOURNAL OF PROTEOME RESEARCH, 2006, 5 (01) :127-134

[10] Analysis of phosphorylation sites on proteins from Saccharomyces cerevisiae by electron transfer dissociation (ETD) mass spectrometry [J].

Chi, An ;

Huttenhower, Curtis ;

Geer, Lewis Y. ;

Coon, Joshua J. ;

Syka, John E. P. ;

Bai, Dina L. ;

Shabanowitz, Jeffrey ;

Burke, Daniel J. ;

Troyanskaya, Olga G. ;

Hunt, Donald F. .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2007, 104 (07) :2193-2198

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