Inhibition of catalytic activity of recombinant mammalian ACs 1, 2, and 5 and bacterial AC toxin CyaA by (M)ANT nucleotides
Activity of ACs 1, 2, and 5 and CyaA was determined in the presence of (M)ANT nucleotides at increasing concentrations. Ki values were calculated by nonlinear regression. Data are the mean values ± S.D. of four to five independent experiments performed in triplicates with at least two different membrane preparations (for mACs).
| (M)ANT Nucleotide | AC 1 | AC 2 | AC 5 | CyaA | |
|---|---|---|---|---|---|
| nM | |||||
| 1 | MANT-ATP | 150 ± 40 | 330 ± 100 | 100 ± 30 | 4300 ± 400 |
| 2 | MANT-ITP | 2.8 ± 0.9 | 13.5 ± 0.5 | 1.2 ± 0.1 | 600 ± 100 |
| 3 | MANT-CTP | 150 ± 30 | 690 ± 20 | 150 ± 30 | 1100 ± 100 |
| 4 | Cl-ANT-ATP | 80 ± 4 | 490 ± 20 | 45 ± 1 | 350 ± 40 |
| 5 | Cl-ANT-ITP | 3.3 ± 0.3 | 8 ± 1 | 2.0 ± 0.2 | 700 ± 200 |
| 6 | Br-ANT-ATP | 120 ± 20 | 450 ± 40 | 70 ± 20 | 330 ± 30 |
| 7 | Br-ANT-ITP | 7.0 ± 0.1 | 22 ± 4 | 4.6 ± 0.4 | 920 ± 50 |
| 8 | Pr-ANT-ATP | 440 ± 20 | 1100 ± 100 | 360 ± 60 | 820 ± 250 |
| 9 | Pr-ANT-ITP | 22 ± 1 | 68 ± 12 | 10 ± 2 | 3800 ± 500 |
| 10 | Ac-NH-ANT-ATP | 6800 ± 200 | 11,000 ± 1000 | 3400 ± 40 | 710 ± 40 |
| 11 | Ac-NH-ANT-ITP | 140 ± 30 | 390 ± 80 | 37 ± 7 | 4800 ± 900 |
| 12 | MANT-ADP | 1300 ± 200 | 2900 ± 500 | 800 ± 200 | 12,000 ± 2000 |
| 13 | MANT-IDP | 39 ± 12 | 86 ± 9 | 31 ± 12 | 11,000 ± 3000 |
| 14 | Br-ANT-ADP | 560 ± 10 | 1700 ± 100 | 280 ± 20 | 8700 ± 1300 |
| 15 | MANT-IMP | 4600 ± 400 | 8200 ± 800 | 3400 ± 200 | >100,000 |
| 16 | ANT-IMP | 7400 ± 1200 | 7500 ± 1,400 | 4300 ± 600 | >100,000 |
| 17 | Bis-MANT-ATP | 700 ± 200 | 2100 ± 600 | 430 ± 50 | 360 ± 10 |
| 18 | Bis-MANT-ITP | 310 ± 20 | 1100 ± 200 | 140 ± 40 | 3000 ± 400 |
| 19 | Bis-MANT-CTP | 620 ± 40 | 7800 ± 100 | 750 ± 40 | 2500 ± 500 |
| 20 | Bis-Cl-ANT-ATP | 1700 ± 100 | 2400 ± 100 | 1600 ± 100 | 16 ± 1 |
| 21 | Bis-Cl-ANT-ITP | 66 ± 1 | 200 ± 10 | 65 ± 3 | 15 ± 1 |
| 22 | Bis-Br-ANT-ATP | 670 ± 50 | 1100 ± 100 | 900 ± 90 | 12.6 ± 0.3 |
| 23 | Bis-Br-ANT-ITP | 21 ± 1 | 71 ± 2 | 15 ± 2 | 20 ± 2 |
| 24 | Bis-Pr-ANT-ATP | 18,000 ± 3000 | 36,000 ± 3000 | 18,000 ± 5000 | 700 ± 200 |
| 25 | Bis-Pr-ANT-ITP | 440 ± 10 | 1400 ± 100 | 250 ± 20 | 2100 ± 100 |
| 26 | Bis-Ac-NH-ANT-ATP | 22,000 ± 1000 | 7000 ± 1000 | 6100 ± 1300 | 280 ± 20 |
| 27 | Bis-Ac-NH-ANT-ITP | 1700 ± 100 | 5600 ± 300 | 480 ± 30 | 7500 ± 100 |
| 28 | Bis-MANT-ADP | 700 ± 300 | 1600 ± 300 | 510 ± 70 | 6500 ± 800 |
| 29 | Bis-MANT-IDP | 1000 ± 100 | 1400 ± 100 | 700 ± 200 | 6600 ± 900 |
| 30 | Bis-Br-ANT-ADP | 1600 ± 300 | 3300 ± 200 | 1800 ± 200 | 91 ± 5 |
| 31 | Bis-MANT-IMP | >100,000 | >100,000 | >100,000 | 20,000 ± 3000 |
| 32 | Bis-ANT-IMP | >100,000 | >100,000 | >100,000 | 12,000 ± 3000 |