[HTML][HTML] A method to identify serine kinase substrates: Akt phosphorylates a novel adipocyte protein with a Rab GTPase-activating protein (GAP) domain
This study describes a method for the identification of the substrates of specific serine
kinases. An antibody specific for the phosphomotif generated by the kinase is used to isolate
phosphorylated substrates by immunoprecipitation, and the isolated proteins are identified
by tandem mass spectrometry of peptides. This method was applied to the identification of
substrates for the protein kinase Akt, which specifically phosphorylates the RXRXXS/T motif.
3T3-L1 adipocytes were treated with insulin to activate Akt, and the putative Akt substrate …
kinases. An antibody specific for the phosphomotif generated by the kinase is used to isolate
phosphorylated substrates by immunoprecipitation, and the isolated proteins are identified
by tandem mass spectrometry of peptides. This method was applied to the identification of
substrates for the protein kinase Akt, which specifically phosphorylates the RXRXXS/T motif.
3T3-L1 adipocytes were treated with insulin to activate Akt, and the putative Akt substrate …