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

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