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

AuthorsCarrascal, Montserrat ; Ovelleiro, David; Casas, Vanessa ; Gay, Marina ; Abián, Joaquín
T lymphocytes
Immune response
Titanium oxide
LymPHOS database
Issue Date6-Nov-2008
PublisherAmerican Chemical Society
CitationJournal of Proteome Research 7(12): 5167–5176 (2008)
AbstractT 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
Appears in Collections:(IIBB) Artículos
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