Figure 3.
The delivery of A-ALP to the vacuole in vps35Δ cells does not require the early endocytic pathway. Wild-type (SNY36–9A), end3-ts (SNY94), vps35Δ (SNY79), end3-ts vps35Δ (LSY9–5B), vps1Δ (SNY38), and end3-ts vps1Δ (SNY96–9D) cells carrying a plasmid expressing A-ALP (pSN55) were analyzed. Each strain was propagated at 22°C for several doublings before being preincubated at 37°C for 10 min, and then pulsed with [35S]methionine/cysteine for 10 min and chased by the addition of unlabeled amino acids. At the indicated times A-ALP was immunoprecipitated from the cultures and analyzed by SDS-PAGE and fluorography. The positions of unprocessed pro-A-ALP (pA-ALP) and mature A-ALP (mA-ALP) are indicated.