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Sorting of the vacuolar enzyme CPY. Wild-type (wt) cells and strains carrying gene deletions as indicated were grown to exponential phase, spheroplasted, labeled for 15 min at 25°C with Tran35S-label, and chased for 30 min at 25°C. The labeled spheroplasts were separated into pellet (intracellular [I]) and supernatant (extracellular [E]) fractions. The presence of CPY in these fractions was determined by SDS-PAGE of these proteins immunoprecipitated with anti-CPY antibodies.
