Table 1. Binding affinity of Scm3 to histone complexes, and equilibrium constants for (Cse4–H4)2 tetramer formation.
| Scm3 | Kdapp (nM) | Hill coefficient | Overall fit (R2) |
|---|---|---|---|
| Cse4–H4 | 17.5 ± 5.2 | 1.1 ± 0.45 | 0.98 |
| Cse4ΔN–H4 | 17.8 ± 2.4 | 1.4 ± 0.23 | 0.98 |
| H3–H4 | 195.8 ± 7.7 | 1.8 ± 0.18 | 0.94 |
| H2A–H2B | 305.0 ± 40.9 | 1.7 ± 0.47 | 0.96 |
| Nap1 | |||
| Cse4–H4 | 2.4 ± 0.33 | 1.3 ± 0.08 | 0.99 |
| H3–H4 | 0.7 ± 0.04 | 1.7 ± 0.15 | 0.99 |
| (Cse4ΔN–H4)2 tetramer formation on DNA from Scm3–Cse4ΔN–H4 | |||
| ‘601’ DNA (147 bp) | 44.1 ± 11.0 | 1.1 ± 0.3 | 0.98 |
| CEN3 DNA (207 bp) | 34.6 ± 10.2 | 1.5 ± 0.6 | 0.96 |
The calculated dissociation constants (Kdapp), Hill coefficients and overall non-linear fit of the data (R2) were calculated from experiments as shown in Figure 1A. Experiments to measure (Cse4ΔN–H4)2 tetramer formation on DNA from a Scm3–Cse4ΔN–H4 trimeric complex are shown in Figure 2C. The apparent Kd for (Cse4ΔN–H4)2 tetramer formation on DNA from a Scm3–Cse4ΔN–H4 trimeric complex was measured using HI-FI. Standard deviations were calculated from at least three replicate experiments.