Figure S6.
Additional Binding Assays, Related to Figures 5 and 6
(A and C) SEC analysis of the indicated species with absorbance at 280 nm. SDS-PAGE were stained with Coomassie Brilliant Blue to visualize all proteins.
(B) Fluorescence polarization experiments were carried out with a synthetic FAMCENP-C1-21 peptide and saturating concentrations of the indicated MIS12C species and polarization monitored at equilibrium (like in Figure 4A). Unlabeled CENP-C1-71 was added as positive control for competition at the indicated competitor concentration. Head2 had no effects as competitor even at high concentrations, suggesting that its intra-molecular interaction with Head1 is of very modest affinity.