FIG. 2.

Ribosome binding analysis of CTSBP2 penta-alanine mutant proteins. (A) Wild-type and mutant forms of CTSBP2 were translated and [³⁵S]Met labeled in rabbit reticulocyte lysate and resolved by 12% SDS-PAGE. (B) The proteins described in the legend for panel A were incubated with purified rat ribosomes and centrifuged through a 20% sucrose cushion. Equal portions (5%) of the supernatant (S) and pellet (P) were resolved by SDS-PAGE and analyzed by phosphorimaging. The luciferase construct was translated and tested as a negative control. (C) The results presented in panel B were quantitated as the percentage of protein in the pellet and normalized relative to the level of pelleting observed for wild-type CTSBP2. The amount of ribosome binding obtained with in vitro-translated hnRNP F (33% pelleting) was considered background and subtracted. The data shown are the averages ± standard errors of results from at least three independent experiments.