%0 Generic %A Witzke, Kathrin E. %A Rosowski, Kristin %A Müller, Christian %A Ahrens, Maike %A Eisenacher, Martin %A Megger, Dominik A. %A Knobloch, Jürgen %A Koch, Andrea %A Bracht, Thilo %A Sitek, Barbara %D 2016 %T Quantitative Secretome Analysis of Activated Jurkat Cells Using Click Chemistry-Based Enrichment of Secreted Glycoproteins %U https://acs.figshare.com/articles/dataset/Quantitative_Secretome_Analysis_of_Activated_Jurkat_Cells_Using_Click_Chemistry-Based_Enrichment_of_Secreted_Glycoproteins/4040046 %R 10.1021/acs.jproteome.6b00575.s001 %2 https://acs.figshare.com/ndownloader/files/6504255 %K Secreted Glycoproteins Quantitative secretome analyses %K proteins cross-verified %K T cell activation %K Marked glycoproteins %K data sets %K affinity purification %K Click Chemistry-Based Enrichment %K serum supplements %K Activated Jurkat Cells %K PXD %K Quantitative Secretome Analysis %K secretome fractions %K biotin method %K 356 proteins %K secretome analyses %K desthiobiotin approach %K bioorthogonal click chemistry %K 59 proteins %K biotin probe %K cell starvation %K cell culture media limit secretome analyses %K Jurkat cells %K serum depletion %K T cells %K pathophysiological changes %K model %X Quantitative secretome analyses are a high-performance tool for the discovery of physiological and pathophysiological changes in cellular processes. However, serum supplements in cell culture media limit secretome analyses, but serum depletion often leads to cell starvation and consequently biased results. To overcome these limiting factors, we investigated a model of T cell activation (Jurkat cells) and performed an approach for the selective enrichment of secreted proteins from conditioned medium utilizing metabolic marking of newly synthesized glycoproteins. Marked glycoproteins were labeled via bioorthogonal click chemistry and isolated by affinity purification. We assessed two labeling compounds conjugated with either biotin or desthiobiotin and the respective secretome fractions. 356 proteins were quantified using the biotin probe and 463 using desthiobiotin. 59 proteins were found differentially abundant (adjusted p-value ≤0.05, absolute fold change ≥1.5) between inactive and activated T cells using the biotin method and 86 using the desthiobiotin approach, with 31 mutual proteins cross-verified by independent experiments. Moreover, we analyzed the cellular proteome of the same model to demonstrate the benefit of secretome analyses and provide comprehensive data sets of both. 336 proteins (61.3%) were quantified exclusively in the secretome. Data are available via ProteomeXchange with identifier PXD004280. %I ACS Publications