The effect of ΔG⁰_vp7,13-mer on RLP assembly efficiency when the initial protein concentration of vp2, vp6 and vp7 is 1.2 M, 7.8 M and 7.8 M, respectively, and the Gibbs free energy of vp2 and vp6 subunit association (ΔG⁰_vp2,3-mer and ΔG⁰_vp6,13-mer, respectively) is equal to −4.08 kcal.mol⁻¹.

<p><b>ΔG⁰_vp7,13-mer</b> varies between −4.08×[0.05 to 1.4] kcal.mol⁻¹.</p

The concentration of assembly intermediates for three initial protein concentration scenarios:

<p>1) <b>[vp2]₀</b> = 1.2 M, <b>[vp6]₀</b> = 7.8 M and <b>[vp7]₀</b> = 7.8 M (stoichiometric ratios – white bars); 2) <b>[vp2]₀</b> = 1.2 M, <b>[vp6]₀</b> = 7.8 M and <b>[vp7]₀</b> = 0.78 M (limitation of vp7 – black bars); 3) <b>[vp2]₀</b> = 12 M, <b>[vp6]₀</b> = 7.8 M and <b>[vp7]₀</b> = 7.8 M (excess of vp2 – grey bars). The Gibbs free energy of vp2, vp6 and vp7 subunit association was assumed to be equal (<b>ΔG⁰_vp2,3-mer</b> = <b>ΔG⁰_vp6,13-mer</b> = <b>ΔG⁰_vp7,13-mer</b> = −4.08 kcal.mol⁻¹).</p

SLP assembly efficiency as function of the initial vp2 concentration, [vp2]₀, for different Gibbs free energy of vp2 subunit association values, ΔG⁰_vp2,3-mer.

<p>The full line represents <b>ΔG⁰_vp2,3-mer</b> = −4.08 kcal.mol⁻¹ as estimated by Zlotnick 1994, the dash lines represent <b>ΔG⁰_vp2,3-mer</b>>−4.08 kcal.mol⁻¹ and the dot lines represent <b>ΔG⁰_vp2,3-mer</b><−4.08 kcal.mol⁻¹. <b>K_d,app</b> represents the pseudo-critical concentration that satisfies the constraint <b>[1]</b> (concentration of unassociated vp2 subunits at equilibrium - <b>[1]</b> = <b>[svp2]_e</b>) = <b>[20]</b> (complete SLP) = <b>K_d,app</b> at equilibrium for a specific <b>ΔG⁰_vp2,3-mer</b>.</p

Kinetics of <i>in vitro</i> RLP assembly and disassembly by 90° light scattering.

<p>Transition of DLP to RLP under standard condition (TNC buffer at pH 5.5, 25°C, NaCl 0.1 M and Ca²⁺ 1 mM) (graph A) and upon increase in pH (8 – graph B), Ca²⁺ (5 mM – graph C), temperature (35°C – graph D) and NaCl (0.5 M – graph E). Dissociation of RLP to DLP at 25°C in TNC with 1 mM of EDTA (grey triangle) or in D-PBS with 1 mM of EGTA (black diamond) (graph F). The arrows indicate the times at which vp7 was added to DLP (graphs A to E) or the addition of chelating agents (graph F).</p

<i>In vitro</i> assembly and disassembly of RLPs.

<p><i>In vitro</i> assembly and disassembly of RLPs.</p