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Closed Access item Phosphorylation analysis of primary human T lymphocytes using sequential IMAC and titanium oxide enrichment

Authors:Carrascal, Montserrat
Ovelleiro, David
Casas, Vanessa
Gay, Marina
Abián, Joaquín
Keywords:Phosphoproteomics, T lymphocytes, Immune response, IMAC, Titanium oxide, LC−MSn, LymPHOS database
Issue Date:6-Nov-2008
Publisher:American Chemical Society
Citation:Journal of Proteome Research 7(12): 5167–5176 (2008)
Abstract: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-MS(n) 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.
Publisher version (URL):http://dx.doi.org/10.1021/pr800500r
E-ISSNmetadata.dc.identifier.doi = DOI:1535-3907
Appears in Collections:(IIBB) Artículos

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