Abstract
Here we demonstrate quantitation of stimuli-induced proteome dynamics in primary cells by combining the power of bio-orthogonal noncanonical amino acid tagging (BONCAT) and stable-isotope labeling of amino acids in cell culture (SILAC). In conjunction with nanoscale liquid chromatography-tandem mass spectrometry (nanoLC-MS/MS), quantitative noncanonical amino acid tagging (QuaNCAT) allowed us to monitor the early expression changes of >600 proteins in primary resting T cells subjected to activation stimuli.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acids
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CD4-Positive T-Lymphocytes / drug effects
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CD4-Positive T-Lymphocytes / metabolism*
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Calcium Ionophores / pharmacology
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Carcinogens / pharmacology
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Chromatography, Liquid / methods
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Gene Expression Regulation / physiology*
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Humans
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Ionomycin / pharmacology
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Isotope Labeling
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Phorbol Esters / pharmacology
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Proteomics / methods*
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Sensitivity and Specificity
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Tandem Mass Spectrometry / methods
Substances
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Amino Acids
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Calcium Ionophores
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Carcinogens
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Phorbol Esters
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Ionomycin