Fig. 1.

ODD-Luc fusion protein retains luciferase activity and binds to pVHL. (A) Autoradiogram of the indicated ³⁵S-labeled proteins after in vitro translation and SDS/polyacrylamide gel electrophoresis. (B) Luciferase activity (LU, light units) corresponding to 50% (by volume) of the in vitro translation products studied in A. Data shown are representative of two independent experiments. (C) Autoradiogram of the indicated ³⁵S-labeled proteins made by in vitro translation in rabbit reticulocyte lysate (Retic) or wheat germ extract (WG). Proteins were incubated with glutathione Sepharose preloaded with GST-pVHL, elongin B, and elongin C. Specifically bound proteins were released by boiling in SDS-containing sample buffer and resolved by SDS-polyacrylamide gel electrophoresis (PD, pulldown). For comparison, 10% of the protein used for the binding studies was load directly and resolved in the same gel (I, input).