Henttu, P.M.A., Kalkhoven, E. and Parker, M.G.
Notes: [35S]methionine-labeled RIP-140, SRC-1, TIF1 and TIF2 were produced using the TNT® Coupled Reticulocyte Lysate System. The labeled proteins were incubated with GST or GST-AF2 receptor fusion proteins coupled to glutathione beads in the absence or presence of oestradiol or 4-hydroxytamoxifen. After overnight incubation and washing away of free protein, bound proteins were extracted and separated by SDS-PAGE. From binding studies with wild type or mutant AF2 receptors, SRC-1, TIF1 and TIF2 require Lys-366 in AF-2 for their interaction, but RIP-140 does not. (1797)