Winter, C. de

tuberin gst -14-3-3␨ beads protein binding hamartin interaction beads -14-3-3␨ 14-3-3␨ lysate variant r 611q variant experiment 14-3-3 phosphorylation phosphatase 4 ° c tuberin-hamartin hela cell lysate yeast antibody erasmus mc 1-tsc isoform two-hybrid 30 ° c tuberin /14-3-3␨ interaction 14-3-3 protein family 10 mm kinase wild-type expression glutathione-sepharose beads tuberin /hamartin binding assay /14-3-3␨ activity fusion wild-type tuberin motif 50 mm edta r 905q variant erasmu presence r 611q substitution 14-3-3- binding motifs 50 mm naf tuberin /143-3␨ binding tuberin variants encoding tuberin /14-3-3␨ binding 14-3-3- domain signal 14-3-3 isoforms 60 min substitution library phosphatase inhibitors protein phosphatase immunoblotting phosphoinositide phosphorylated target incubation 14-3-3- binding sites yeast two-hybrid system yeast two-hybrid assay yeast two-hybrid experiments 6 kinase activity r 905q variants /hamartin gbd-tuberin ⁄14-3-3␨ dominant-negative akt isoform overexpressing co-immunoprecipitation experiments akt-mediated phosphorylation www.jbc.org r 905q