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Bioinformatic prediction and confirmation of beta-adducin as a novel substrate of glycogen synthase kinase 3.
Farghaian, Hovik; Turnley, Ann M; Sutherland, Calum; Cole, Adam R.
Affiliation
  • Farghaian H; Neurosignalling Group, Garvan Institute of Medical Research, Sydney, New South Wales 2010, Australia.
J Biol Chem ; 286(28): 25274-83, 2011 Jul 15.
Article de En | MEDLINE | ID: mdl-21606488
ABSTRACT
It is important to identify the true substrates of protein kinases because this illuminates the primary function of any kinase. Here, we used bioinformatics and biochemical validation to identify novel brain substrates of the Ser/Thr kinase glycogen synthase kinase 3 (GSK3). Briefly, sequence databases were searched for proteins containing a conserved GSK3 phosphorylation consensus sequence ((S/T)PXX(S/T)P or (S/T)PXXX(S/T)P), as well as other criteria of interest (e.g. brain proteins). Importantly, candidates were highlighted if they had previously been reported to be phosphorylated at these sites by large-scale phosphoproteomic studies. These criteria identified the brain-enriched cytoskeleton-associated protein ß-adducin as a likely substrate of GSK3. To confirm this experimentally, it was cloned and subjected to a combination of cell culture and in vitro kinase assays that demonstrated direct phosphorylation by GSK3 in vitro and in cells. Phosphosites were mapped to three separate regions near the C terminus and confirmed using phosphospecific antibodies. Prior priming phosphorylation by Cdk5 enhanced phosphorylation by GSK3. Expression of wild type, but not non-phosphorylatable (GSK3 insensitive), ß-adducin increased axon and dendrite elongation in primary cortical neurons. Therefore, phosphorylation of ß-adducin by GSK3 promotes efficient neurite outgrowth in neurons.
Sujet(s)

Texte intégral: 1 Collection: 01-internacional Base de données: MEDLINE Sujet principal: Axones / Cortex cérébral / Protéines du cytosquelette / Dendrites / Glycogen Synthase Kinase 3 Type d'étude: Prognostic_studies / Risk_factors_studies Limites: Humans Langue: En Journal: J Biol Chem Année: 2011 Type de document: Article Pays d'affiliation: Australie

Texte intégral: 1 Collection: 01-internacional Base de données: MEDLINE Sujet principal: Axones / Cortex cérébral / Protéines du cytosquelette / Dendrites / Glycogen Synthase Kinase 3 Type d'étude: Prognostic_studies / Risk_factors_studies Limites: Humans Langue: En Journal: J Biol Chem Année: 2011 Type de document: Article Pays d'affiliation: Australie