TABLE 1.
Summary of kinetic data for the interaction of wild-type and mutant Ad5 fiber knob proteins with sCAR, as determined by SPRa
| Protein assayed | ka (Ms−1) (% WT) | kd (s−1) (% WT) | Kd (nM) |
|---|---|---|---|
| Wild type | (1.8 ± 0.02) × 105 | (2.8 ± 0.06) × 10−3 | 14.8 ± 4.5 |
| Ala406Lys | (2.4 ± 0.20) × 105 (133) | (1.5 ± 0.20) × 10−2 (535) | 73.7 ± 9.8 |
| Arg412Asp | (5.0 ± 0.11) × 104 (27) | (1.5 ± 0.40) × 10−2 (535) | 227 ± 77 |
| Asn414Lys | (2.0 ± 0.07) × 105 (111) | (3.7 ± 0.50) × 10−3 (132) | 19.2 ± 2.8 |
| His456Glu | (1.4 ± 0.03) × 105 (78) | (2.6 ± 0.10) × 10−3 (93) | 17.8 ± 0.9 |
| Ile458Glu | (1.2 ± 0.01) × 105 (67) | (1.7 ± 0.06) × 10−3 (60) | 14.3 ± 5.5 |
| Asn470Glu | (1.6 ± 0.03) × 105 (89) | (3.6 ± 0.04) × 10−3 (129) | 21.5 ± 2.4 |
| Phe472Lys | (2.1 ± 0.17) × 105 (117) | (2.7 ± 0.07) × 10−3 (96) | 12.7 ± 1.2 |
| Arg481Glu | (8.9 ± 1.80) × 104 (49) | (1.2 ± 0.12) × 10−2 (429) | 137 ± 4.5 |
| Asn482Glu | (1.3 ± 0.01) × 105 (72) | (2.3 ± 0.04) × 10−3 (82) | 17.9 ± 1.2 |
| Gly483Glu | (1.4 ± 0.01) × 105 (78) | (5.4 ± 0.02) × 10−3 (192) | 37.8 ± 1.9 |
| Asp484Ala | (1.6 ± 0.04) × 105 (89) | (3.6 ± 0.01) × 10−3 (128) | 21.7 ± 3.8 |
| Lys506Ala | (1.4 ± 0.02) × 105 (78) | (2.9 ± 0.02) × 10−3 (103) | 20.7 ± 1.4 |
| Ser507Ala | (1.8 ± 0.02) × 105 (100) | (2.9 ± 0.01) × 10−3 (105) | 16.1 ± 2.1 |
| His508Glu | (1.6 ± 0.05) × 105 (89) | (4.2 ± 0.18) × 10−3 (150) | 27.2 ± 1.5 |
| Lys510Glu+Lys506Ala | (1.2 ± 0.01) × 105 (67) | (3.2 ± 0.26) × 10−3 (114) | 26.1 ± 2.6 |
a
Sensorgrams were analyzed using a monophasic model for interaction. Results are expressed as mean ± sd for six separate determinations.