Title : Phosphorylation of gephyrin in hippocampal neurons by cyclin-dependent kinase CDK5 at Ser-270 is dependent on collybistin - Kuhse_2012_J.Biol.Chem_287_30952 |
Author(s) : Kuhse J , Kalbouneh H , Schlicksupp A , Mukusch S , Nawrotzki R , Kirsch J |
Ref : Journal of Biological Chemistry , 287 :30952 , 2012 |
Abstract :
Gephyrin is a scaffold protein essential for the postsynaptic clustering of inhibitory glycine and different subtypes of GABA(A) receptors. The cellular and molecular mechanisms involved in gephyrin-mediated receptor clustering are still not well understood. Here we provide evidence that the gephyrin-binding protein collybistin is involved in regulating the phosphorylation of gephyrin. We demonstrate that the widely used monoclonal antibody mAb7a is a phospho-specific antibody that allows the cellular and biochemical analysis of gephyrin phosphorylation at Ser-270. In addition, another neighbored epitope determinant was identified at position Thr-276. Analysis of the double mutant gephyrin(T276A,S277A) revealed significant reduction in gephyrin cluster formation and altered oligomerization behavior of gephyrin. Moreover, pharmacological inhibition of cyclin-dependent kinases in hippocampal neurons reduced postsynaptic gephyrin mAb7a immunoreactivities. In vitro phosphorylation assays and phosphopeptide competition experiments revealed a phosphorylation at Ser-270 depending on enzyme activities of cyclin-dependent kinases CDK1, -2, or -5. These data indicate that collybistin and cyclin-dependent kinases are involved in regulating the phosphorylation of gephyrin at postsynaptic membrane specializations. |
PubMedSearch : Kuhse_2012_J.Biol.Chem_287_30952 |
PubMedID: 22778260 |
Kuhse J, Kalbouneh H, Schlicksupp A, Mukusch S, Nawrotzki R, Kirsch J (2012)
Phosphorylation of gephyrin in hippocampal neurons by cyclin-dependent kinase CDK5 at Ser-270 is dependent on collybistin
Journal of Biological Chemistry
287 :30952
Kuhse J, Kalbouneh H, Schlicksupp A, Mukusch S, Nawrotzki R, Kirsch J (2012)
Journal of Biological Chemistry
287 :30952