Figure 3.

Protease digestion of in vitro translated PPARα. Rat PPARα was in vitro translated and labeled with S³⁵ methionine. Unincorporated S³⁵ was removed with size exclusion resin and the labeled protein was concentrated. Yield was determined and 10 fmol of PPARα were used in each digestion. PPARα was bound to the ligand for 30 minutes at 22°C and then digested with 50 μg/mL of α-chymotrypsin for 20 minutes at 22°C. Resulting digests were resolved on an 18% tris-glycine gel. The constant product is shown with the dark arrow and the Wy-14,643-protected fragments are shown with light arrows, representative of 3 independent experiments.