Structure of the vacuolar ATPase by electron microscopy.
The structure of the vacuolar ATPase from bovine brain clathrin-coated vesicles has been determined by electron microscopy of negatively stained, detergent-solubilized enzyme molecules. Preparations of both lipid-containing and delipidated enzyme have been analyzed. The complex is organized in two major domains, a V(1) and V(0), with overall dimensions of 28 x 14 x 14 nm. The V(1) is a more or less spherical molecule with a central cavity. The V(0) has the shape of a flattened sphere or doughnut with a radius of about 100 A. The V(1) and V(0) are joined by a 60-A long and 40-A wide central stalk, consisting of several individual protein densities. Two kinds of smaller densities are visible at the top periphery of the V(1), and one of these seems to extend all the way down to the stalk domain in some averages. Images of both the lipid-containing and the delipidated complex show a 30-50-kDa protein density on the lumenal side of the complex, opposite the central stalk, centered in the ring of c subunits. A large trans-membrane mass, probably the C-terminal domain of the 100-kDa subunit a, is seen at the periphery of the c subunit ring in some projections. This large mass has both a lumenal and a cytosolic domain, and it is the cytosolic domain that interacts with the central stalk. Two to three additional protein densities can be seen in the V(1)-V(0) interface, all connected to the central stalk. Overall, the structure of the V-ATPase is similar to the structure of the related F(1)F(0)-ATP synthase, confirming their common origin.
Wilkens
S
S
Department of Biochemistry, University of California, Riverside, California 92521, USA.
Vasilyeva
E
E
Forgac
M
M
D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D003034:Coated Pits, Cell-Membrane / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D008854:Microscopy, Electron; D017494:Proton Pumps / Q000648:ultrastructure*; D006180:Proton-Translocating ATPases / Q000648:ultrastructure*; D025262:Vacuolar Proton-Translocating ATPases
D017494:Proton Pumps; D025262:Vacuolar Proton-Translocating ATPases; D006180:Proton-Translocating ATPases