Table 1.
Binding Parameters Determined for Msh4-Msh5 DNA Interactions
| DNA Substrate | Dissociation Constant Kd (nM) |
Protein/DNA Stoichiometry | |
|---|---|---|---|
| EMSAa | Fluorescence Intensityb | ||
| Duplex | 120 ± 33 | 40 ± 15 | 1:1 |
| 3′ overhangs | 47 ± 9 | 15 ± 4 | 1:1 |
| ss forks | 25 ± 7 | 5 ± 1 | 1:1 |
| DLIS | 12 ± 2 | 5 ± 2.5 | 1:1 |
| Pre-HJ | 15 ± 5 | 6 ± 1 | 1:1 |
| Open HJ | 88 ± 22 | 46 ± 7 | 1:1 |
| HJ (I) | 12 ± 2 | 0.3 ± 0.1 | 1:1 |
| HJ (R) | 14 ± 2 | 0.3 ± 0.1 | 1:1 |
a
EMSA Kd values are determined from the fraction of free DNA.
b
Kd values determined from the change in fluorescence intensity or anisotropy as a function of Msh4-Msh5 concentration, as described in the Materials and Methods. All data shown are an average from at least three independent measurements.