Figure 6.

prAPI is protease accessible in the apg5^ts mutant. Spheroplasts isolated from apg5^ts pep4Δ and ypt7Δ cells were shifted to 38°C for 5 min, labeled for 5 min, and then chased for 60 min. A portion of the sample was removed and immunoprecipitated as a total (T) control. After lysis in PS200 with 5 mM MgCl₂ and centrifugation at 5000 × g, supernatant (S) and pellet (P) fractions were collected and subjected to protease treatment, as indicated. The resulting fractions were subsequently immunoprecipitated with antiserum against API, CPY, and hexokinase (cytosolic marker). Hexokinase was recovered 78 and 81% in the supernatant fraction in apg5^ts pep4Δ and ypt7Δ cells, respectively, indicating efficient lysis of the spheroplasts. The graph represents the percentage of prAPI and p2CPY recovered in the pellet fraction that is protease resistant (lane 4 divided by lane 3). TX-100, Triton X-100.