title: Glucose-regulated and drug-perturbed phosphoproteome reveals molecular mechanisms controlling insulin secretion.
abstract: Insulin-secreting beta cells play an essential role in maintaining physiological blood glucose levels, and their dysfunction leads to the development of diabetes. To elucidate the signalling events regulating insulin secretion, we applied a recently developed phosphoproteomics workflow. We quantified the time-resolved phosphoproteome of murine pancreatic cells following their exposure to glucose and in combination with small molecule compounds that promote insulin secretion. The quantitative phosphoproteome of 30,000 sites clustered into three main groups in concordance with the modulation of the three key kinases: PKA, PKC and CK2A. A high-resolution time course revealed key novel regulatory sites, revealing the importance of methyltransferase DNMT3A phosphorylation in the glucose response. Remarkably a significant proportion of these novel regulatory sites is significantly downregulated in diabetic islets. Control of insulin secretion is embedded in an unexpectedly broad and complex range of cellular functions, which are perturbed by drugs in multiple ways.
authors.affiliation: Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried 82152, Germany.
authors.forename: Francesca
authors.identifier: 0000-0001-5586-9529
authors.initials: F
authors.lastname: Sacco
authors.affiliation: Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried 82152, Germany.
authors.forename: Sean J
authors.initials: SJ
authors.lastname: Humphrey
authors.affiliation: Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried 82152, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Cox
authors.affiliation: Sanofi Aventis Deutschland GmbH, R&D, LGCR, SDI, Bioinformatics, Frankfurt 65926, Germany.
authors.forename: Marcel
authors.initials: M
authors.lastname: Mischnik
authors.affiliation: Sanofi Aventis Deutschland GmbH, Global Diabetes Division, R&TM, Islet Biology, Frankfurt 65926, Germany.
authors.forename: Anke
authors.initials: A
authors.lastname: Schulte
authors.affiliation: Sanofi Aventis Deutschland GmbH, R&D, LGCR, SDI, Bioinformatics, Frankfurt 65926, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Klabunde
authors.affiliation: Sanofi Aventis Deutschland GmbH, Global Diabetes Division, R&TM, Islet Biology, Frankfurt 65926, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Schäfer
authors.affiliation: Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried 82152, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Mann
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D016000:Cluster Analysis; D003922:Diabetes Mellitus, Type 1 / Q000378:metabolism; D005947:Glucose / Q000494:pharmacology*; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000187:drug effects* / Q000378:metabolism; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D010750:Phosphoproteins / Q000145:classification / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D054852:Small Molecule Libraries / Q000494:pharmacology*
chemical_list: D007328:Insulin; D010750:Phosphoproteins; D020543:Proteome; D054852:Small Molecule Libraries; D005947:Glucose