Figure 6.
The vps35–101 and vps35–108 alleles exhibit specific defects in A-ALP and Vps10p trafficking, respectively. The VPS35, vps35–101, vps35–108, and vps35Δ yeast strains correspond to the following strain/plasmid combinations, respectively: SNY79/pSN54/pLS13, SNY79/pSN54/pLS12, SNY79/pSN54/pPB7–010.4, and PBY3/pSN54 (please refer to Tables 1 and 2 for strain and plasmid information). (A) The strains were pulsed for 10 min, chased for the indicated times, and subjected to A-ALP immunoprecipitation as detailed in the legend of Figure 4. (B) The strains were pulsed with [35S]methionine/cysteine for 10 min, and then chased for 45 min with unlabeled amino acids. CPY was immunoprecipitated from intracellular (I) and extracellular (E) fractions and analyzed by SDS-PAGE and fluorography. The positions of precursor (proCPY) and mature (mCPY) forms of CPY are indicated.