title: Experimental and computational tools for analysis of signaling networks in primary cells.
abstract: Cellular information processing in signaling networks forms the basis of responses to environmental stimuli. At any given time, cells receive multiple simultaneous input cues, which are processed and integrated to determine cellular responses such as migration, proliferation, apoptosis, or differentiation. Protein phosphorylation events play a major role in this process and are often involved in fundamental biological and cellular processes such as protein-protein interactions, enzyme activity, and immune responses. Determining which kinases phosphorylate specific phospho sites poses a challenge; this information is critical when trying to elucidate key proteins involved in specific cellular responses. Here, methods to generate high-quality quantitative phosphorylation data from cell lysates originating from primary cells, and how to analyze the generated data to construct quantitative signaling network models, are presented. These models can subsequently be used to guide follow-up in vitro/in vivo validation studies.
authors: Cellular Signal Integration Group (C-SIG), Center for Biological Sequence Analysis (CBS), Department of Systems Biology, Technical University of Denmark (DTU), Lyngby, Denmark.
authors: Erwin M
authors: EM
authors: Schoof
authors: Cellular Signal Integration Group (C-SIG), Center for Biological Sequence Analysis (CBS), Department of Systems Biology, Technical University of Denmark (DTU), Lyngby, Denmark.
authors: Rune
authors: R
authors: Linding
mesh_terms: D000818:Animals; D002460:Cell Line; D019295:Computational Biology / Q000379:methods*; D003198:Computer Simulation; D006801:Humans; D013058:Mass Spectrometry; D053858:Metabolic Networks and Pathways; D010766:Phosphorylation; D061251:Primary Cell Culture; D011485:Protein Binding; D015398:Signal Transduction; D012984:Software
keywords: mass spectrometry; network biology; phosphorylation; primary cell signaling