Figure S7.
Comparison of BKA-Inhibited M-State Structures from Crystals Grown with and without Nanobody, Related to Figure 6
(A) Superposition of m-state carrier structures from crystals grown with nanobody (P3221 space group, gray tube) and without nanobody (P212121 space group, tube with domain 1, blue; domain 2, yellow; domain 3, red), viewed from the membrane. The structures superimpose with 0.6 Å RMSD over 1096 atoms, showing that presence of the nanobody, different detergent and crystallization conditions, and different crystal packings have not perturbed the structure of the BKA-inhibited state. (B) The same superposition as (A), viewed from the cytoplasmic side of the membrane. (C) Stereo-view of electron density maps for the P212121 crystal form, following molecular replacement (using individual domains from the P3221 crystal) and rigid-body refinement in AutoBuster. The P3221 structure is shown as a gray cartoon, with the P212121 colored by domain. (D) Electron density for domain 1. (E) Partial density for BKA, not included in the molecular replacement search models, gives confidence in the quality of the molecular replacement phases. In (C)-(E), the 2mFo-DFc electron density map, contoured at 1 times the root mean square electron density (σ), is shown as a blue mesh, and the mFo-DFc electron density map is shown contoured at +3σ (green mesh) and −3σ (red mesh).