Structure of Na+,K+-ATPase at 11-A resolution: comparison with Ca2+-ATPase in E1 and E2 states.
Na+,K+-ATPase is a heterodimer of alpha and beta subunits and a member of the P-type ATPase family of ion pumps. Here we present an 11-A structure of the heterodimer determined from electron micrographs of unstained frozen-hydrated tubular crystals. For this reconstruction, the enzyme was isolated from supraorbital glands of salt-adapted ducks and was crystallized within the native membranes. Crystallization conditions fixed Na+,K+-ATPase in the vanadate-inhibited E2 conformation, and the crystals had p1 symmetry. A large number of helical symmetries were observed, so a three-dimensional structure was calculated by averaging both Fourier-Bessel coefficients and real-space structures of data from the different symmetries. The resulting structure clearly reveals cytoplasmic, transmembrane, and extracellular regions of the molecule with densities separately attributable to alpha and beta subunits. The overall shape bears a remarkable resemblance to the E2 structure of rabbit sarcoplasmic reticulum Ca2+-ATPase. After aligning these two structures, atomic coordinates for Ca2+-ATPase were fit to Na+,K+-ATPase, and several flexible surface loops, which fit the map poorly, were associated with sequences that differ in the two pumps. Nevertheless, cytoplasmic domains were very similarly arranged, suggesting that the E2-to-E1 conformational change postulated for Ca2+-ATPase probably applies to Na+,K+-ATPase as well as other P-type ATPases.
Rice
W J
WJ
Skirball Institute of Biomolecular Medicine and Department of Cell Biology, New York University Medical Center, New York, New York 10016, USA.
Young
H S
HS
Martin
D W
DW
Sachs
J R
JR
Stokes
D L
DL
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D000252:Calcium-Transporting ATPases / Q000737:chemistry*; D002462:Cell Membrane / Q000737:chemistry; D018360:Crystallography, X-Ray; D019281:Dimerization; D004372:Ducks; D004583:Electrons; D005583:Fourier Analysis; D007091:Image Processing, Computer-Assisted; D007477:Ions; D008854:Microscopy, Electron; D008958:Models, Molecular; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000201:enzymology; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry*
D007477:Ions; D000251:Adenosine Triphosphatases; D000252:Calcium-Transporting ATPases; D000254:Sodium-Potassium-Exchanging ATPase