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Figure 6.
Fig. 6. Inhibition of in vitro assembly by NYAD-1. (a)
Negatively stained EM images of immature- and mature-like
particles resulting from in vitro assembly of Gag (25 μM) and
CA proteins (50 μM), respectively, in the presence of no
peptide (a and g, control), 0.25-fold (b and h), 0.5-fold (c and
i), a molar equivalent (d and j), and fivefold molar equivalent
of NYAD-1 (e and k) and CAI (f and l). A dose-response effect
was observed with NYAD-1. The integrity of the mature-like
particles is shown in the insets. Gag in vitro assembly was
conducted by dialyzing against 50 mM Na[2]HPO[4], pH 8.0
containing 0.1 M NaCl in the presence of 5% total E. coli RNA
(RNA/protein = 1:20, w/w). The CA assembly reaction was
initiated at a final concentration of 1.2 M NaCl. (b)
Dosage-dependent inhibition of Gag assembly; 20 fields under EM
were screened and the number of VLPs was plotted against the
ratio of concentration of NYAD-1 to Gag proteins in the assembly
reaction.
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