FIG. 5.

In vitro translation of protease constructs (A₃₄₈/G₁₃₀₁, M₈₂₇/G₁₃₀₁, V₉₂₀/G₁₃₀₁, A₉₇₄/G₁₃₀₁, A₁₀₂₀/G₁₃₀₁, and G₁₁₀₂/G₁₃₀₁) and examination of their trans-cleavage activities. In vitro transcription and translation were carried out as described in Materials and Methods. A₃₄₈/G₁₃₀₁ (A), M₈₂₇/G₁₃₀₁ (B), V₉₂₀/G₁₃₀₁ (C), A₉₇₄/G₁₃₀₁ (D), A₁₀₂₀/G₁₃₀₁ (E), and G₁₁₀₂/G₁₃₀₁ (F) were translated individually to give 102-, 50-, 41-, 35-, 30-, and 21-kDa products, respectively (lanes 1). Cotranslation of each construct with substrate p200(C1152S) was carried out at 30°C for 0 to 5 h. Samples were removed at each time point and subjected to SDS-PAGE analysis (lanes 2 to 7). Positions of molecular mass markers and cleavage products are indicated. Images were scanned using a UMAX Astra 1220U scanner with Adobe Photoshop 5.0 software.