Fig. 2: Characterization of BGL homotrimers.

a Cartoon representation of six representative successful redesigns of BGL0, with two from each design set: Helix Rebuilding plus C-terminal attachment (HR-C), Helix Rebuilding plus N-terminal attachment (HR-N), and Normal Modes (NM) relaxation. Blue and orange colored backbones highlight N- and C-terminal two-helix segments comprising the interface, which was redesigned, and newly designed hydrogen bond networks (HBNets) are shown in sticks with the hydrogen bonds in dashed lines. Design names in the bottom left corner of each box and box outline and trace colors are used consistently throughout. Characterization of the six examples from (a) by (b) SEC, (c) SAXS, (d) nsEM, and (e) nMS. b Overlay of SEC traces normalized to maximum absorbance at 280 nm (A280), after soluble aggregate removal and TEV cleavage. SEC traces of all 20 designs are provided in Supplementary Fig. 4. c Overlay of SAXS data. Data were scaled appropriately to align at q = 0.01 to highlight the similarity of the profiles. d Individual 2D class averages of nsEM data for each sample. e Individual mass deconvolved nMS data of TEV-cleaved samples (Supplementary Data 13). All peaks that match the expected mass in Daltons for each homotrimer are annotated with green circles, and homohexamers (likely to be dimers of trimers) are annotated with orange diamonds.