1. Fig. 1 Data collection at a 40° tilt angle enabled reconstruction of a cryo-EM map of the NRTN-GFRα2-RET complex at a 5.7-Å resolution.
Data collection at a 40° tilt angle enabled reconstruction of a cryo-EM map of the NRTN-GFRα2-RET complex at a 5.7-Å resolution. (A) Comparison of two-dimensional (2D) class averages obtained from data collected at 0° or 40° tilt angles. 2D classes from the 0° data collection showed severe orientation bias of the molecule, exclusively presenting the view from the top revealing a figure 8 shape. 2D classes from data collected at the 40° tilt angle show a broader distribution of views. (B) Cryo-EM map reconstruction at a 5.7-Å resolution and atomic model derived from fitting in the cryo-EM map. Density around the NRTN dimer (or cartoon representation) is colored orange, GFRα2 is in blue, and RETCLD1-4 is in pink and magenta. Density for the unmodeled RETCRD is depicted in gray. Most of the density could be modeled, and only RETCRD could not be traced (gray, center). The side view of the complex suggests the position of the plasma membrane (right). (C) Schematic close-up view of the calcium-binding site between RETCLD2 and RETCLD3 (left), GFRα2 domains 1–3 showing the electron density (center), and the NRTN-GFRα2 interface mediated by the NRTN finger domain (right).
Janna M. Bigalke et al. Sci Adv 2019;5:eaau4202
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