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 assayedka (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.