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Table 1 The effect of metal ions on the kinetic parameters of HIV-1 sIN binding to DNA substrates

From: Mode of inhibition of HIV-1 Integrase by a C-terminal domain-specific monoclonal antibody*

 

U5-End

U5-End Cut

Target

 

kon 105 M-1 s-1

koff 10-3 s-1

Kd (nM)

kon 105 M-1 s-1

koff 10-3 s-1

Kd (nM)

kon 105 M-1 s-1

koff 10-3 s-1

Kd (nM)

sIN

1.7 ± 0.1

1.0 ± 0.1

5.9 ± 0.7

1.1 ± 0.1

1.3 ± 0.1

11.8 ± 1.4

1.0 ± 0.1

1.6 ± 0.2

16.0 ± 2.6

sIN no Mg

1.9 ± 0.2

2.1 ± 0.1

11.1 ± 1.3

2.7 ± 0.2

2.0 ± 0.1

7.4 ± 0.7

3.5 ± 0.2

2.1 ± 0.1

6.0 ± 0.7

  1. The model U5 end biotin-labeled substrate (see Experimental Procedures) was immobilized on a SA chip. BIAcore data were collected by injecting different concentrations of sIN, ranging from 50 to 300 nM dimers, at a flow rate of 30 μL/min. No Mg2+ ions were achieved by eliminating MgCl2 from the running buffer and including 0.5 mM EDTA in the running as well as the dilution buffer. The dissociation constant was calculated from the ratio of the average of the off and the apparent on rates. All injections were done at 25°C.