Figure 2: Recombinant TAF2–8–10 complex.

(a) TAF2, the TAF8–10 pair and a mixture of TAF2–8–10 were analysed by SEC. Elution profiles of TAF2 (green), TAF8–10 (blue) and TAF2–8–10 (purple) are plotted in relative absorption units at 280 nm versus elution volume (top). Fractions are numbered (top of graph). SDS–PAGE analyses of the eluted samples are shown (below). Molecular masses of protein standards are indicated on the left of gel sections. Protein denominations are shown on the right. First lane shows the SEC input (IN). (b) Absorbance c(s) profiles from sedimentation velocity analytical ultracentrifugation experiments are plotted for TAF2 (green), TAF8–10 (blue) and TAF2–8–10 (purple). (c) Mass spectrum of TAF2–8–10 complex electrosprayed from an aqueous ammonium acetate solution under high collision energy for subunit dissociation. The MS spectrum reveals peaks with corresponding masses for a TAF8–10 dimer (blue dots), TAF2 subunit (green dots) and a predominant TAF2–8–10 complex (purple dots) centred at 4,000, 6,000 and 7,500 m/z, respectively. At 12,000 m/z is a TAF2–8 dimer (yellow dots) resulting from the dissociation of the TAF10 subunit (light blue dots) from the intact TAF2–8–10 complex. Proteins and protein complexes are shown schematically as coloured circles.