Figure 2. Electron crystallography provides structures of aquaporin-0 (AQP0) in sphingomyelin/ cholesterol bilayers at 2.5 Å resolution.
(A) Electron diffraction pattern of an untilted AQP0 two-dimensional (2D) crystal reconstituted at a sphingomyelin:cholesterol ratio of 2:1, showing reflections to ~2 Å resolution. Scale bar indicates (10 Å)–1. (B) Density map at 2.5 Å resolution used to build the AQP02SM:1Chol structure. A region of the water-conducting pathway close to the NPA (asparagine-proline-alanine), the AQP signature motif, is shown. The 2Fo-Fc map contoured at 1.5σ is shown as gray mesh, the AQP0 model is shown in yellow with oxygen atoms in red and nitrogen atoms in blue. The red sphere represents a water molecule. (C) A diffraction pattern of an untilted AQP0 2D crystal reconstituted at a sphingomyelin:cholesterol ratio of 1:2, showing reflections to better than 1.6 Å resolution. Scale bar indicates (10 Å)–1. (D) Density map at 2.5 Å resolution used to build the AQP01SM:2Chol structure. The same region as in (B) is shown with the same color code. See also Figure 2—figure supplement 1.

