2-D structure of the Neurospora crassa plasma membrane ATPase as determined by electron cryomicroscopy.
Large, well-ordered 2-D crystals of the dodecylmaltoside complex of the Neurospora crassa plasma membrane H(+)-ATPase grow rapidly on the surface of a polyethylene glycol-containing mixture similar to that originally developed for growing 3-D crystals of this integral membrane transport protein. Negative stain electron microscopy of the crystals shows that many are single layers. Cryoelectron microscopy of unstained specimens indicates that the crystals have a p6 layer group with unit cell dimensions of a = b = 167 A. Image processing of selected electron micrographs has yielded a projection map at 10.3 A resolution. The repeating unit of the ATPase crystals comprises six 100 kDa ATPase monomers arranged in a symmetrical ring. The individual monomers in projection are shaped like a boot. These results provide the first indications of the molecular structure of the H(+)-ATPase molecule. They also establish the feasibility of precipitant-induced surface growth as a rapid, simple alternative to conventional methods for obtaining 2-D crystals of the integral membrane proteins useful for structure analysis.
Cyrklaff
M
M
European Molecular Biology Laboratory, Heidelberg, Germany.
Auer
M
M
Kühlbrandt
W
W
Scarborough
G A
GA
D002462:Cell Membrane / Q000201:enzymology; D003460:Crystallization; D005583:Fourier Analysis; D007091:Image Processing, Computer-Assisted; D008854:Microscopy, Electron; D015394:Molecular Structure; D009492:Neurospora crassa / Q000201:enzymology*; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000648:ultrastructure*
D006180:Proton-Translocating ATPases