Table 1.
Competition | Fluorescence | ||||||
---|---|---|---|---|---|---|---|
Analog | Kd (nM) | Fold Loss | ΔΔGbind (kcal/mol) |
Kd (nM) | Fold Loss | ΔΔGbind (kcal/mol) |
|
c-di-GMP | 1.4 ± 0.1a | - | - | 16b | - | - | |
| |||||||
Base | c-GMP-IMP | 39 ± 2.4 | 27 | 2.0 | 75 ± 14 | 5 | 0.9 |
c-di-IMP | 1100 ± 210 | 790 | 3.9 | ≥ 15,000 | ≥ 940 | ≥ 4.1 | |
c-c7G-GMP | 1100 ± 80 | 790 | 3.9 | ≥ 15,000 | ≥ 940 | ≥ 4.1 | |
c-di-c7GMP | n.b.c | - | - | n.b. | - | - | |
c-GMP-AMP | 1600 ± 160 | 1200 | 4.0 | - | - | - | |
c-di-AMP | n.b. | - | - | n.b. | - | - | |
c-N1mG-GMP | 420 ± 16 | 300 | 3.4 | - | - | - | |
| |||||||
Ribose | c-dG-GMP | 51 ± 5.4 | 37 | 2.1 | 140 ± 24 | 9 | 1.3 |
c-di-dGMP | 2600 ± 88 | 1800 | 4.4 | ≥ 20,000 | ≥ 1300 | ≥ 4.2 | |
c-2’F-G-GMP | - | - | - | 25 ± 3.7 | 2 | 0.3 | |
c-di-2’F-GMP | 56 ± 9.0 | 40 | 2.2 | 180 ± 15 | 11 | 1.4 | |
c-2’OMe-G-GMP | 200 ± 31 | 150 | 2.9 | - | - | - | |
c-di-2’OMe-GMP | ≥ 30,000 | ≥ 21,000 | 5.9 | n.b. | - | - | |
| |||||||
Phos. | c-(Rp)-Gps-GMP | - | - | - | 67 ± 11 | 4 | 0.8 |
c-(Sp)-Gps-GMP | - | - | - | 280 ± 40 | 18 | 1.7 | |
c-(RpRp)-di-Gps | 150 ± 33 | 100 | 2.8 | 320 ± 70 | 20 | 1.8 | |
c-(RpSp)-di-Gps | 750 ± 63 | 540 | 3.7 | 1700 ± 160 | 110 | 2.8 |
Measured by direct binding.
Value determined by Kd = koff/kon (see supplemental info, Table S2).
No detectable binding at highest ligand concentration tested (250 μM by competition, 20 μM by fluorescence).