Figure 3.
CD mutant G129E preserves protein phosphatase activity but lacks inositol phosphatase activity and is incapable of inducing a G1 block. (A) Comparison of wild-type PTEN and the mutants G129R and G129E in the cell-cycle assay. 786-O cells were cotransfected with pCD19 and plasmids encoding the indicated proteins and analyzed as in Fig. 1A. (B) Comparison of the expression of wild-type PTEN and the mutants G129R and G129E in 786-O cells. 786-O cells were transfected with plasmids encoding the indicated proteins. Anti-HA immunoprecipitates of protein extracts prepared from metabolically labeled cells were separated by gel electrophoresis and subject to fluorography. (C) Inositol phosphatase activity of GST-PTEN and mutant derivatives. The indicated GST-PTEN fusion proteins were analyzed as in Fig. 2C. (D) Protein tyrosine phosphatase activity of GST-PTEN and mutant derivatives. The indicated GST-PTEN fusion proteins were analyzed as in Fig. 2D. (A, C, and D) The mean and SEM of two experiments are shown.