Table 2.
RNA molecule | Contacts affected by modification | Modified phosphate oxygen |
Kd, nM
|
Specificity | |
---|---|---|---|---|---|
U-RNA | C-RNA | ||||
RNAs 5 and 6 | |||||
Unmodified | 10, 1.7* | 0.04* | 146 | ||
Deoxy −5 | Glu-63 | 270, 23* | 1.8* | 81 | |
RNAs 7 and 8 | |||||
Unmodified | 13, 4.1* | 0.04* | 214 | ||
RP-thioP-3 | None | O1P | 19, 4.8* | 0.04* | 298 |
SP-thioP-3 | None | O2P | 9.4, 5.7* | 0.03* | 252 |
RP-thioP-7 | Lys-57 | O1P | 2.7, 1.4* | 0.05* | 41 |
SP-thioP-7 | Asn-55, Ser-52 | O2P | 51, 15* | 1.3* | 25 |
SP-methylP-7 | Lys-57 | O1P | 88, 16* | 26* | 2.0 |
RP-methylP-7 | Asn-55, Ser-52 | O2P | 295, 34* | 18* | 9.1 |
RP-thioP-6 | Cytosine-5 | O1P | 0.6, 0.2* | 0.02* | 20 |
SP-thioP-6 | None | O2P | 7.9, 2.1* | 0.03* | 167 |
DithioP-6 | Cytosine-5 | O1P, O2P | 0.4, 0.06* | 0.04* | 5.8 |
SP-methylP-6 | Cytosine-5 | O1P | 17, 6.2* | 23* | 0.5 |
RP-methylP-6 | None | O2P | 34, 9.2* | 3.3* | 6.5 |
Kd values were measured in 80 mM KCl/10 mM MgCl2/100 mM K–Hepes, pH 7.5, at 0°C. Specificity is defined as Kd (U-RNA)/Kd (C-RNA) and was calculated by using an average of the kinetic and thermodynamic determinations of the U-RNA.
Calculated by dividing the experimental off-rate by 108 M−1⋅min−1.