Nm 23 H1 co immunoprecipitated KSR from lysates of transiently transfected 293T cells and at endogenous protein expression levels in MDA MB 435 breast carcinoma cells . ^^^ Autophosphorylated recombinant Nm 23 H1 phosphorylated KSR in vitro . ^^^ Phosphoamino acid analysis identified serine as the major target , and two peaks of Nm 23 H1 phosphorylation were identified upon high performance liquid chromatography analysis of KSR tryptic peptides . ^^^ Using site directed mutagenesis , we found that Nm 23 H1 phosphorylated KSR serine 392 , a 14 3 3 binding site , as well as serine 434 when serine 392 was mutated . ^^^ |