Extended Data Fig. 1: High-resolution ssNMR studies of the teixobactin - Lipid II complex in membranes.

All ssNMR spectra were acquired with uniformly ¹³C,¹⁵N-labelled teixobactin bound to ¹²C,¹⁴N-Lipid II in DOPC liposomes. a) 2D ¹³C¹³C PARISxy³³ spectrum acquired at 950 MHz and 18 kHz MAS using 50 ms CC magnetization transfer time. All ¹³C nuclei of teixobactin are assigned. b) 2D NH spectrum acquired at 700 MHz and 60 kHz MAS. All signals are assigned. c) 2D CN spectrum acquired at 700 MHz and 12 kHz MAS using 5 ms N to C cross-polarization time. Some connectivities, including for the End10 sidechain, are shown by dashed lines. Note that our 2D NH data of the ¹³C,¹⁵N-teixobactin – Lipid II complex in membranes match well to a previously published spectrum of the complex in dodecylphosphocholine (DPC) micelles²². Interestingly, compared to micellar assignments, we observe major (>5 ¹³C ppm) local differences for the two residues that most prominently interact with Lipid II, something that could suggest an impact of the membrane environment for complex formation. See Supplementary Table 3 for a comparison of chemical shifts of the teixobactin – Lipid II complex in micelles and in membranes. Note that backbone assignments were conclusively validated with a pair of ¹H-detected 3D CαNH/CONH experiments⁶⁵.