Large-Scale Functional Purification of Recombinant HIV-1 Capsid
Figure 5
Inhibition of full-length wild-type capsid polymerization kinetics by two CA binding agents.
(A, B) CAI-4 is a peptide that binds to the C-terminal domain of CA. (C, D) BM2 is a small molecule inhibitor described by Boehringer Ingelheim that binds to the CA N-terminal domain. WT CA was pre-incubated with each inhibitor. Assembly reactions were induced by 2-fold dilution of capsid protein into buffer containing 50 mM sodium phosphate pH 7.5, 4 M NaCl at 25°C. The increase of turbidity (absorbance at 350 nm) of 40 µM solution of WT CA in the presence of increasing concentrations of inhibitor (depicted on the right side of kinetic traces) was plotted as a function of time. The data points represent an average of three determinations with standard deviation shown as grey bars. The dose response curves of inhibition of polymerization kinetics and the concentration of inhibitor resulting in a half-maximal inhibition (IC50) for (B) CAI-4 and (D) BM2. The percent of inhibitor activity (% Activity) represents the percent of decrease of the initial rate of polymerization reaction.