Figure 6.
Glo3-R59K causes defects in CPY maturation and secretion of the ER-resident protein Kar2. (A) Proliferating cells were exposed to [35S]methionine and cysteine for 7 min and sampled without a chase with unlabeled amino acids at 0, 30, and 60 min. CPY was immunoprecipitated, resolved by SDS-PAGE, and detected by autoradiography. This panel is representative of three independent experiments. (B) Expression of MET3pr-glo3-R59K, MET3pr-GLO3, or MET3pr-empty vector was induced by incubation in medium lacking methionine for 6 h at 30°C. The medium was collected, proteins were precipitated, and analyzed by immunoblot. Kar2 was secreted only by cells expressing the ArfGAP-dead Glo3-R59K.