Figure 7.

In vivo degradation of heat-denatured Hsp150Δ–β-lactamase. (A) H621 (lhs1⁻ sec18), (B) H647 (lhs1⁻ c-myc–LHS1 ⁺ sec18), and (C) H534 (Hsp104⁻ sec18) cells were preincubated for 15 min at 37°C, labeled in SC medium lacking methionine and cysteine with [³⁵S]methionine/cysteine for 20 min, diluted with excess SC medium containing unlabeled methionine and cysteine, and incubated further at 37°C for 40 min (lanes 1). Parallel cells were incubated thereafter for 20 min at 50°C (lanes 2), or after this at 24°C for 2 h (lanes 3), 4 h (lanes 4), 6 h (lanes 5), or 8 h (lanes 6). The cells were lysed and immunoprecipitated with anti– β-lactamase antiserum, followed by SDS-PAGE analysis.