Structure determination of clathrin coats to subnanometer resolution by single particle cryo-electron microscopy.
Clathrin triskelions can assemble into lattices of different shapes, sizes and symmetries. For many years, the structures of clathrin lattices have been studied by single particle cryo-electron microscopy, which probed the architecture of the D6 hexagonal barrel clathrin coat at the molecular level. By introducing additional image processing steps we have recently produced a density map for the D6 barrel clathrin coat at subnanometer resolution, enabling us to generate an atomic model for this lattice [Fotin, A., Cheng, Y., Sliz, P., Grigorieff, N., Harrison, S.C., Kirchhausen, T., Walz, T., 2004. Molecular model for a complete clathrin lattice from electron cryomicroscopy. Nature 432, 573-579]. We describe in detail here the image processing steps that we have added to produce a density map at this high resolution. These procedures should be generally applicable and may thus help determine the structures of other large protein assemblies to higher resolution by single particle cryo-electron microscopy.
Fotin
Alexander
A
Biophysics Graduate Program, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.
Kirchhausen
Tomas
T
Grigorieff
Nikolaus
N
Harrison
Stephen C
SC
Walz
Thomas
T
Cheng
Yifan
Y
D002966:Clathrin / Q000737:chemistry*; D022163:Clathrin-Coated Vesicles / Q000737:chemistry*; D020285:Cryoelectron Microscopy / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008958:Models, Molecular
D002966:Clathrin