Searching for novel Cdk5 substrates in brain by comparative phosphoproteomics of wild type and Cdk5-/- mice.
Searching for novel Cdk5 substrates in brain by comparative phosphoproteomics of wild type and Cdk5-/- mice.
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DOI：
10.1371/journal.pone.0090363
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
González-Billault C
中科院分区：
文献类型：
--
作者：
Contreras-Vallejos E;Utreras E;Bórquez DA;Prochazkova M;Terse A;Jaffe H;Toledo A;Arruti C;Pant HC;Kulkarni AB;González-Billault C
Protein phosphorylation is the most common post-translational modification that regulates several pivotal functions in cells. Cyclin-dependent kinase 5 (Cdk5) is a proline-directed serine/threonine kinase which is mostly active in the nervous system. It regulates several biological processes such as neuronal migration, cytoskeletal dynamics, axonal guidance and synaptic plasticity among others. In search for novel substrates of Cdk5 in the brain we performed quantitative phosphoproteomics analysis, isolating phosphoproteins from whole brain derived from E18.5 Cdk5+/+ and Cdk5−/− embryos, using an Immobilized Metal-Ion Affinity Chromatography (IMAC), which specifically binds to phosphorylated proteins. The isolated phosphoproteins were eluted and isotopically labeled for relative and absolute quantitation (iTRAQ) and mass spectrometry identification. We found 40 proteins that showed decreased phosphorylation at Cdk5−/− brains. In addition, out of these 40 hypophosphorylated proteins we characterized two proteins, :MARCKS (Myristoylated Alanine-Rich protein Kinase C substrate) and Grin1 (G protein regulated inducer of neurite outgrowth 1). MARCKS is known to be phosphorylated by Cdk5 in chick neural cells while Grin1 has not been reported to be phosphorylated by Cdk5. When these proteins were overexpressed in N2A neuroblastoma cell line along with p35, serine phosphorylation in their Cdk5 motifs was found to be increased. In contrast, treatments with roscovitine, the Cdk5 inhibitor, resulted in an opposite effect on serine phosphorylation in N2A cells and primary hippocampal neurons transfected with MARCKS. In summary, the results presented here identify Grin 1 as novel Cdk5 substrate and confirm previously identified MARCKS as a a bona fide Cdk5 substrate.
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影响因子：
64.5
作者：
Huttlin EL;Jedrychowski MP;Elias JE;Goswami T;Rad R;Beausoleil SA;Villén J;Haas W;Sowa ME;Gygi SP
通讯作者：
Gygi SP
影响因子：
3.4
作者：
Goswami, Tapasree;Li, Xue;Smith, Aidan M.;Luderowski, Eva M.;Vincent, James J.;Rush, John;Ballif, Bryan A.
通讯作者：
Ballif, Bryan A.
影响因子：
4.8
作者：
Chen, LT;Gilman, AG;Kozasa, T
通讯作者：
Kozasa, T
影响因子：
9.2
作者：
Connell-Crowley, L;Le Gall, M;Giniger, E
通讯作者：
Giniger, E
影响因子：
2.1
作者：
Horiuchi, Youko;Asada, Akiko;Nishizawa, Masafumi
通讯作者：
Nishizawa, Masafumi