Extended Data Fig. 1: Immature VLPs assemble at 100-fold lower IP6 concentrations than mature capsids.
From: HIV-1 is dependent on its immature lattice to recruit IP6 for mature capsid assembly
(A) In vitro mature assembly kinetics with 75 µM CA and 50–1500 µM IP6. (B) Maximum assembly from (A) at different IP6 concentrations fit to equation I: Y = Amin + (Xh)*(Amax-Amin)/(EC50MAh + Xh); where Amin and Amax is the minimum and maximum assembly, EC50MA is the effective concentration for half-maximal mature assembly and h is the Hill slope. Fitting gave an EC50MA of 250 ± 40 µM. Error bars depict mean Amax ± SD of N = 3 independent experiments. (C) EM images of negatively stained samples of the final assembly reaction from (A) using 1.5 mM IP6. Size bars are 200 nm. (D) In vitro immature VLP assembly with 75 µM ΔMA-CANC and a range of IP6 concentrations. (E) Data from (D) were fitted as in (B) to give an EC50IA of 3 ± 1 µM. Error bars depict mean Amax ± SD of N = 3 independent experiments. (F) EM images of negatively stained samples of the final assembly reaction from (C) using 14 µM IP6. Size bars are 100 nM.
