Figure 5: Fractal assemblies capture and release greater amounts of cargo compared to globular assemblies

Figure 5: Fractal assemblies capture and release greater amounts of cargo compared to globular assemblies. Scheme for the envisioned reversible capture of cargo proteins for the (A) fractal and (B) globular structures. The red stars denote an example cargo protein (antibody). (C) % protein capture was measured for 3:2 fractal (assemblies obtained with 3 AtzA-pY: 2 AtzC-SH2), 3:2 GS linker (globular assemblies obtained at the same stoichiometry with fusion proteins containing long GS-rich linkers), and 3:1 fractal (assemblies obtained with 3 AtzA-pY: 1 AtzC-SH2). The 3:2 fractal captured more IgG antibody (red bars) and GFP-SH2 (green barss), and degraded more substrate TCP (purple bar; reflecting the higher capture efficiency of enzyme DhaA-Sh2) than the 3:2 GS linker assemblies. In addition, 3:2 fractal released more captured antibody compared to the 3:2 GS linker when incubated with YopH phosphatase (blue bars). (D, E) Confocal fluorescence microscopy images of the 3-component assembly with GFP-SH2 showing the topology of incorporation of GFP-SH2 in fractal and (F-G) the incorporation of GFP-SH2 into the globular assemblies. (H-I) the IgG antibody Alexa Fluor 568 incorporation into the fractal assembly and (J-K) the incorporation into the globular assembly.