title: Alterations in the cerebellar (Phospho)proteome of a cyclic guanosine monophosphate (cGMP)-dependent protein kinase knockout mouse.
abstract: The cyclic nucleotide cyclic guanosine monophosphate (cGMP) plays an important role in learning and memory, but its signaling mechanisms in the mammalian brain are not fully understood. Using mass-spectrometry-based proteomics, we evaluated how the cerebellum adapts its (phospho)proteome in a knockout mouse model of cGMP-dependent protein kinase type I (cGKI). Our data reveal that a small subset of proteins in the cerebellum (∼3% of the quantified proteins) became substantially differentially expressed in the absence of cGKI. More changes were observed at the phosphoproteome level, with hundreds of sites being differentially phosphorylated between wild-type and knockout cerebellum. Most of these phosphorylated sites do not represent known cGKI substrates. An integrative computational network analysis of the data indicated that the differentially expressed proteins and proteins harboring differentially phosphorylated sites largely belong to a tight network in the Purkinje cells of the cerebellum involving important cGMP/cAMP signaling nodes (e.g. PDE5 and PKARIIβ) and Ca(2+) signaling (e.g. SERCA3). In this way, removal of cGKI could be linked to impaired cerebellar long-term depression at Purkinje cell synapses. In addition, we were able to identify a set of novel putative (phospho)proteins to be considered in this network. Overall, our data improve our understanding of cerebellar cGKI signaling and suggest novel players in cGKI-regulated synaptic plasticity.
authors.affiliation: From ‡Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands; §Netherlands Proteomics Centre, Padualaan 8, 3584 CH Utrecht, The Netherlands;
authors.forename: Eleonora
authors.initials: E
authors.lastname: Corradini
authors.affiliation: ¶Interfakultäres Institut für Biochemie, Universität Tübingen, D-72074 Tübingen, Germany; ‖Graduate School for Cellular and Molecular Neuroscience, Universität Tübingen, D-72074 Tübingen, Germany; **German Center for Neurodegenerative diseases (DZNE), D-72076 Tübingen, Germany.
authors.forename: Raghavan
authors.initials: R
authors.lastname: Vallur
authors.affiliation: From ‡Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands; §Netherlands Proteomics Centre, Padualaan 8, 3584 CH Utrecht, The Netherlands;
authors.forename: Linsey M
authors.initials: LM
authors.lastname: Raaijmakers
authors.affiliation: ¶Interfakultäres Institut für Biochemie, Universität Tübingen, D-72074 Tübingen, Germany;
authors.forename: Susanne
authors.initials: S
authors.lastname: Feil
authors.affiliation: ¶Interfakultäres Institut für Biochemie, Universität Tübingen, D-72074 Tübingen, Germany;
authors.forename: Robert
authors.initials: R
authors.lastname: Feil
authors.affiliation: From ‡Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands; §Netherlands Proteomics Centre, Padualaan 8, 3584 CH Utrecht, The Netherlands; a.j.r.heck@uu.nl a.scholten@uu.nl.
authors.forename: Albert J R
authors.initials: AJ
authors.lastname: Heck
authors.affiliation: From ‡Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands; §Netherlands Proteomics Centre, Padualaan 8, 3584 CH Utrecht, The Netherlands; a.j.r.heck@uu.nl a.scholten@uu.nl.
authors.forename: Arjen
authors.initials: A
authors.lastname: Scholten
mesh_terms: D000818:Animals; D002531:Cerebellum / Q000378:metabolism*; D006152:Cyclic GMP; D062965:Cyclic GMP-Dependent Protein Kinase Type I / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D010750:Phosphoproteins / Q000302:isolation & purification*; D010766:Phosphorylation; D040901:Proteomics / Q000379:methods; D015398:Signal Transduction; D013569:Synapses / Q000378:metabolism
chemical_list: D010750:Phosphoproteins; D062965:Cyclic GMP-Dependent Protein Kinase Type I; C569720:Prkg1 protein, mouse; D006152:Cyclic GMP