5 research outputs found
The concentration of assembly intermediates for three initial protein concentration scenarios:
<p>1) <b>[vp2]<sub>0</sub></b> = 1.2 M, <b>[vp6]<sub>0</sub></b> = 7.8 M and <b>[vp7]<sub>0</sub></b> = 7.8 M (stoichiometric ratios – white bars); 2) <b>[vp2]<sub>0</sub></b> = 1.2 M, <b>[vp6]<sub>0</sub></b> = 7.8 M and <b>[vp7]<sub>0</sub></b> = 0.78 M (limitation of vp7 – black bars); 3) <b>[vp2]<sub>0</sub></b> = 12 M, <b>[vp6]<sub>0</sub></b> = 7.8 M and <b>[vp7]<sub>0</sub></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>ΔG<sup>0</sup><sub>vp2,3-mer</sub></b> = <b>ΔG<sup>0</sup><sub>vp6,13-mer</sub></b> = <b>ΔG<sup>0</sup><sub>vp7,13-mer</sub></b> = −4.08 kcal.mol<sup>−1</sup>).</p
The effect of ΔG<sup>0</sup><sub>vp7,13-mer</sub> 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 (ΔG<sup>0</sup><sub>vp2,3-mer</sub> and ΔG<sup>0</sup><sub>vp6,13-mer</sub>, respectively) is equal to −4.08 kcal.mol<sup>−1</sup>.
<p><b>ΔG<sup>0</sup><sub>vp7,13-mer</sub></b> varies between −4.08×[0.05 to 1.4] kcal.mol<sup>−1</sup>.</p
SLP assembly efficiency as function of the initial vp2 concentration, [vp2]<sub>0</sub>, for different Gibbs free energy of vp2 subunit association values, ΔG<sup>0</sup><sub>vp2,3-mer</sub>.
<p>The full line represents <b>ΔG<sup>0</sup><sub>vp2,3-mer</sub></b> = −4.08 kcal.mol<sup>−1</sup> as estimated by Zlotnick 1994, the dash lines represent <b>ΔG<sup>0</sup><sub>vp2,3-mer</sub></b>>−4.08 kcal.mol<sup>−1</sup> and the dot lines represent <b>ΔG<sup>0</sup><sub>vp2,3-mer</sub></b><−4.08 kcal.mol<sup>−1</sup>. <b>K<sub>d,app</sub></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]<sub>e</sub></b>) = <b>[20]</b> (complete SLP) = <b>K<sub>d,app</sub></b> at equilibrium for a specific <b>ΔG<sup>0</sup><sub>vp2,3-mer</sub></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 Ca<sup>2+</sup> 1 mM) (graph A) and upon increase in pH (8 – graph B), Ca<sup>2+</sup> (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