Extended Data Fig. 7: Crystal structure of the domain-swapped DHD_15 and biophysical characterization of higher-order oligomers. | Nature

Extended Data Fig. 7: Crystal structure of the domain-swapped DHD_15 and biophysical characterization of higher-order oligomers.

From: Programmable design of orthogonal protein heterodimers

Extended Data Fig. 7

a, Crystal structure of DHD_15 at pH 6.5, with 2.25 Å resolution. b, Superposition of design models (in colour) onto both halves of the crystal structure (in white), with backbone r.m.s.d. of 1.83 Å. c, Native MS study of DHD_15 at different pH values indicates that heterodimers, rather than heterotetramers, are dominant in solution. dg, SEC traces of the induced dimerization DHD_9-13 fusion (d), DHD_15-37 fusion (e), DHD_13-37 fusion (f), and the scaffolding complex in Fig. 3d (g; the peak at around 15 ml corresponds to the fully assembled complex, followed by a peak representing an excess of individual components). h, CD thermal melt curves for the scaffolding complex in Fig. 3d. Wavelength scan was performed at 25 °C, 75 °C, 95 °C and final 25 °C. Design was α-helical and stable up to 95 °C. i, CD chemical denaturation profile of the scaffolding complex in Fig. 3d. Two (cg) or one (h, i) biologically independent repeats were performed.