Fig. 1: Comparisons of tES–F116H folding.
From: A general approach to protein folding using thermostable exoshells
a Description of the tES nanoencapsulation protocol depicting buffer adjustments and buffer exchange via dialysis (b) Crude soluble yields of POI increase after tES–F116H nanoencapsulation. As the amount of tES added to the folding mix was the limiting factor for POI-soluble yield, the results are expressed as mg POI/100 mg of tES–F116H. c Fold increase of soluble POI folded with tES–F116H compared with its absence. d Functional yields of POI are determined by the ratio of POI to charge complementary tES–F116Hsubunits in the folding mix and demonstrate saturation with titration. e Charge complementation of the tES–F116H interior with POI determines stabilizing energetics. Three tES–F116H charge variants are assayed and POI self-organizes according to their charge (labeled next to the ordinate) and stabilizing effect. f Low-molecular-weight proteins require high molar ratios to stabilize the assembly. Thermal denaturation temperatures of tES–F116H were measured via DFS as the concentation of POI is titrated against tES–F116H during the loading phase of the folding protocol. g tES–F116H undergo pH-mediated assembly and disassembly. Shell diameter can be monitored using DLS (lower panel) and no significant soluble protein loss is observed after 10 cycles (upper panel). (b–g, Data are presented as mean ± SEM., n = 3 independent experiments) (Source data are provided as a Source Data file).
