Mapping PP6c binding by charge-reversal mutations in PP6R3. (A) FLAG-tagged full length (FL) PP6R3 (A), and mutants E63-64K (B), D113R (C), E204-205K (D), E259-262K (E) and E204-205-259-262K (F) were expressed in HEK293 cells and immunoprecipitated using immobilized anti-FLAG antibody. Co-precipitated PP6c was quantified by fluorescent immunoblotting and normalized for the amount of FLAG-tagged protein. Results were replicated in 3 independent experiments and plotted as mean +/- SD. (B) Immunoblot of co-precipitated endogenous PP6c from one experiment (upper panel) and the FLAG-tagged PP6R3 proteins (lower panel).
Guergnon et al. BMC Biochemistry 2009 10:24 doi:10.1186/1471-2091-10-24