PMID- 25907765 OWN - NLM STAT- MEDLINE DCOM- 20160502 LR - 20211203 IS - 1535-9484 (Electronic) IS - 1535-9476 (Print) IS - 1535-9476 (Linking) VI - 14 IP - 8 DP - 2015 Aug TI - Functional Proteomics Identifies Acinus L as a Direct Insulin- and Amino Acid-Dependent Mammalian Target of Rapamycin Complex 1 (mTORC1) Substrate. PG - 2042-55 LID - 10.1074/mcp.M114.045807 [doi] AB - The serine/threonine kinase mammalian target of rapamycin (mTOR) governs growth, metabolism, and aging in response to insulin and amino acids (aa), and is often activated in metabolic disorders and cancer. Much is known about the regulatory signaling network that encompasses mTOR, but surprisingly few direct mTOR substrates have been established to date. To tackle this gap in our knowledge, we took advantage of a combined quantitative phosphoproteomic and interactomic strategy. We analyzed the insulin- and aa-responsive phosphoproteome upon inhibition of the mTOR complex 1 (mTORC1) component raptor, and investigated in parallel the interactome of endogenous mTOR. By overlaying these two datasets, we identified acinus L as a potential novel mTORC1 target. We confirmed acinus L as a direct mTORC1 substrate by co-immunoprecipitation and MS-enhanced kinase assays. Our study delineates a triple proteomics strategy of combined phosphoproteomics, interactomics, and MS-enhanced kinase assays for the de novo-identification of mTOR network components, and provides a rich source of potential novel mTOR interactors and targets for future investigation. CI - (c) 2015 by The American Society for Biochemistry and Molecular Biology, Inc. FAU - Schwarz, Jennifer Jasmin AU - Schwarz JJ AD - From the double daggerFaculty of Biology, Institute of Biology III, University of Freiburg, 79104 Freiburg, Germany; section signFaculty of Biology, Institute of Biology II, University of Freiburg, 79104 Freiburg, Germany; paragraph signSpemann Graduate School of Biology and Medicine (SGBM), University of Freiburg; FAU - Wiese, Heike AU - Wiese H AD - section signFaculty of Biology, Institute of Biology II, University of Freiburg, 79104 Freiburg, Germany; FAU - Tolle, Regine Charlotte AU - Tolle RC AD - From the double daggerFaculty of Biology, Institute of Biology III, University of Freiburg, 79104 Freiburg, Germany; section signFaculty of Biology, Institute of Biology II, University of Freiburg, 79104 Freiburg, Germany; ||Department of Dermatology, Medical Center, and Centre for Biological Systems Analysis (ZBSA), University of Freiburg, 79104 Freiburg, Germany; FAU - Zarei, Mostafa AU - Zarei M AD - ||Department of Dermatology, Medical Center, and Centre for Biological Systems Analysis (ZBSA), University of Freiburg, 79104 Freiburg, Germany; **BIOSS Centre for Biological Signaling Studies, University of Freiburg, 79104 Freiburg, Germany; FAU - Dengjel, Jorn AU - Dengjel J AD - paragraph signSpemann Graduate School of Biology and Medicine (SGBM), University of Freiburg; ||Department of Dermatology, Medical Center, and Centre for Biological Systems Analysis (ZBSA), University of Freiburg, 79104 Freiburg, Germany; **BIOSS Centre for Biological Signaling Studies, University of Freiburg, 79104 Freiburg, Germany; FAU - Warscheid, Bettina AU - Warscheid B AD - section signFaculty of Biology, Institute of Biology II, University of Freiburg, 79104 Freiburg, Germany; paragraph signSpemann Graduate School of Biology and Medicine (SGBM), University of Freiburg; **BIOSS Centre for Biological Signaling Studies, University of Freiburg, 79104 Freiburg, Germany; k.thedieck@umcg.nl kathrin.thedieck@uni-oldenburg.de bettina.warscheid@biologie.uni-freiburg.de. FAU - Thedieck, Kathrin AU - Thedieck K AD - From the double daggerFaculty of Biology, Institute of Biology III, University of Freiburg, 79104 Freiburg, Germany; **BIOSS Centre for Biological Signaling Studies, University of Freiburg, 79104 Freiburg, Germany; double daggerdouble daggerDepartment of Pediatrics, University of Groningen, University Medical Center Groningen (UMCG), 9713 AV Groningen, The Netherlands; section sign section signDepartment for Neurosciences, Faculty VI - School of Medicine and Health Sciences, Carl von Ossietzky University Oldenburg, 26129 Oldenburg, Germany k.thedieck@umcg.nl kathrin.thedieck@uni-oldenburg.de bettina.warscheid@biologie.uni-freiburg.de. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150423 PL - United States TA - Mol Cell Proteomics JT - Molecular & cellular proteomics : MCP JID - 101125647 RN - 0 (ACIN1 protein, human) RN - 0 (Adaptor Proteins, Signal Transducing) RN - 0 (Amino Acids) RN - 0 (Insulin) RN - 0 (Multiprotein Complexes) RN - 0 (Nuclear Proteins) RN - 0 (Phosphoproteins) RN - 0 (Proteome) RN - 0 (RPTOR protein, human) RN - 0 (Regulatory-Associated Protein of mTOR) RN - EC 2.7.11.1 (Mechanistic Target of Rapamycin Complex 1) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) SB - IM MH - Adaptor Proteins, Signal Transducing/metabolism MH - Amino Acid Sequence MH - Amino Acids/*metabolism MH - Gene Knockdown Techniques MH - HeLa Cells MH - Humans MH - Insulin/*metabolism MH - Mechanistic Target of Rapamycin Complex 1 MH - Molecular Sequence Data MH - Multiprotein Complexes/*metabolism MH - Nuclear Proteins/chemistry/*metabolism MH - Phosphoproteins/metabolism MH - Phosphorylation MH - Protein Interaction Mapping MH - Proteome/metabolism MH - Proteomics/*methods MH - Regulatory-Associated Protein of mTOR MH - Substrate Specificity MH - TOR Serine-Threonine Kinases/*metabolism PMC - PMC4528236 EDAT- 2015/04/25 06:00 MHDA- 2016/05/03 06:00 PMCR- 2016/08/01 CRDT- 2015/04/25 06:00 PHST- 2014/10/29 00:00 [received] PHST- 2015/04/25 06:00 [entrez] PHST- 2015/04/25 06:00 [pubmed] PHST- 2016/05/03 06:00 [medline] PHST- 2016/08/01 00:00 [pmc-release] AID - S1535-9476(20)32659-1 [pii] AID - M114.045807 [pii] AID - 10.1074/mcp.M114.045807 [doi] PST - ppublish SO - Mol Cell Proteomics. 2015 Aug;14(8):2042-55. doi: 10.1074/mcp.M114.045807. Epub 2015 Apr 23.