Figure 5

Half-lives of wild-type, Rab27a^T23N, Rab27a^Q78L and Rab27a^N133I mutant proteins. COS-7 cells transfected with either pCAGGS myc-Rab27a^WT (A), pCAGGS myc-Rab27a^T23N (B), pCAGGS myc-Rab27a^Q78L (C), or co-transfected with pCAGGS myc-Rab27a^WT and pCAGGS myc-Rab27a^T23N (D) were labelled with [³⁵S]methionine/cysteine for 2 h and subsequently chased in Dulbecco's modified Eagle's medium for the indicated times. Rab27a protein was immunoprecipitated from cell extracts, collected with protein G-Sepharose beads, separated on a 12% SDS-polyacrylamide gel and autoradiographed as described under "Methods". (Panel E) Quantification of radiolabelled samples obtained above.