Sievert, HenningHenningSievertVenz, SimoneSimoneVenzPlatas Barradas, OscarOscarPlatas BarradasDhople, Vishnu MukundVishnu MukundDhopleSchaletzky, MartinMartinSchaletzkyNagel, Claus-HenningClaus-HenningNagelBraig, MelanieMelanieBraigPreukschas, MichaelMichaelPreukschasPällmann, NoraNoraPällmannBokemeyer, CarstenCarstenBokemeyerBrümmendorf, Tim H.Tim H.BrümmendorfPörtner, RalfRalfPörtnerWalther, ReinhardReinhardWaltherDuncan, Kent E.Kent E.DuncanHauber, JoachimJoachimHauberBalabanov, StefanStefanBalabanov2019-01-212019-01-212012-08-10Molecular & cellular proteomics : MCP 11 (11): 1289-1305 (2012)https://tubdok.tub.tuhh.de/handle/11420/1940Hypusine modification of eukaryotic initiation factor 5A (eIF-5A) represents a unique and highly specific post-translational modification with regulatory functions in cancer, diabetes, and infectious diseases. However, the specific cellular pathways that are influenced by the hypusine modification remain largely unknown. To globally characterize eIF-5A and hypusine-dependent pathways, we used an approach that combines large-scale bioreactor cell culture with tandem affinity purification and mass spectrometry: "bioreactor-TAP-MS/MS." By applying this approach systematically to all four components of the hypusine modification system (eIF-5A1, eIF-5A2, DHS, and DOHH), we identified 248 interacting proteins as components of the cellular hypusine network, with diverse functions including regulation of translation, mRNA processing, DNA replication, and cell cycle regulation. Network analysis of this data set enabled us to provide a comprehensive overview of the protein-protein interaction landscape of the hypusine modification system. In addition, we validated the interaction of eIF-5A with some of the newly identified associated proteins in more detail. Our analysis has revealed numerous novel interactions, and thus provides a valuable resource for understanding how this crucial homeostatic signaling pathway affects different cellular functions.en1535-948420121112891305American Society for Biochemistry and Molecular Biology, HighWire Presshttps://creativecommons.org/licenses/by/4.0/AnimalsComputational BiologyDNA-Binding ProteinsHumansLysineMass SpectrometryMiceMixed Function OxygenasesMultivesicular BodiesNIH 3T3 CellsNuclear ProteinsOxidoreductases Acting on CH-NH Group DonorsPeptide FragmentsPeptide Initiation FactorsProtein TransportRNA-Binding ProteinsRecombinant Fusion ProteinsReproducibility of ResultsRibosomal ProteinsSubcellular FractionsProtein Interaction MapsProtein Processing, Post-TranslationalBiowissenschaften, BiologieMedizinJournal Articleurn:nbn:de:gbv:830-882.02499410.15480/882.193711420/194010.1074/mcp.M112.01905910.15480/882.1937Other