TABLE 2.
Dissociation constants for oligonucleotide binding to recombinant proteins and complexes
The column labeled “Protein” indicates the recombinant protein or protein complex used for assays of binding to the column header DNA oligonucleotides. Numbers for Teb1, TEB, Rpa1, RPA, Rlp1, and RTT are given in nm as calculated from three experimental replicates of the gel mobility shift assays. S.E. was calculated for each mean to give an estimate of the variation among the replicates. All values had p < 0.06 for goodness of fit for the one-site binding model used to calculate the dissociation constant. −, binding affinity too low to quantify using the gel mobility shift assay conditions. Rows labeled “-Fold change” indicate the relative increase in binding comparing the preceding large subunit protein alone to the heterotrimer.
| Protein | (GT2G3)3 | dT18 | (GT2G3)5 | dT30 |
|---|---|---|---|---|
| Teb2-Teb3 | − | − | − | − |
| Teb1 (nm) | 3.4 ± 1.3 | − | 2.5 ± 1.0 | − |
| TEB (nm) | 2.9 ± 0.8 | − | 2.2 ± 1.7 | − |
| -Fold change | 1.2 | − | 1.1 | − |
| Rpa1 (nm) | 36 ± 11 | 41 ± 20 | 36 ± 15 | 39 ± 16 |
| RPA (nm) | 24 ± 9 | 30 ± 12 | 7.4 ± 3.0 | 5.3 ± 0.6 |
| -Fold change | 1.5 | 1.4 | 4.8 | 7.3 |
| Rlp1 (nm) | 68 ± 22 | 75 ± 9 | 73 ± 28 | 81 ± 14 |
| RTT (nm) | 45 ± 16 | 56 ± 25 | 27 ± 13 | 21 ± 7 |
| -Fold change | 1.5 | 1.3 | 2.7 | 3.8 |