Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1987 Jul 1;105(1):1–8. doi: 10.1083/jcb.105.1.1

The three-dimensional structure of the Na,K-ATPase from electron microscopy

PMCID: PMC2114893  PMID: 3038922

Abstract

The structure of Na,K-ATPase has been studied by electron microscopy and image reconstruction. A three-dimensional structure of this enzyme has been obtained to an overall resolution of 2.5 nm using data from specimens of negatively stained dimer sheets tilted through a range of angles +/- 60 degrees. The reconstruction shows a complex mass distribution consisting of ribbons of paired molecules extending approximately 6.0 nm from the cytoplasmic side of the membrane. The molecular envelope consists of a massive "body" with "lobe" and "arm" structures projecting from it. The body has a columnar shape and is tilted with respect to the plane of the membrane. The region of interaction responsible for dimer formation is located between two bodies and is clearly visible in the reconstruction. It has been identified as a segment in the amino-terminal portion of the alpha subunit. The arms that interconnect the ribbons are located close to the membrane and are most probably formed by the beta subunits.

Full Text

The Full Text of this article is available as a PDF (1.9 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aebi U., Fowler W. E., Isenberg G., Pollard T. D., Smith P. R. Crystalline actin sheets: their structure and polymorphism. J Cell Biol. 1981 Nov;91(2 Pt 1):340–351. doi: 10.1083/jcb.91.2.340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brisson A., Unwin P. N. Tubular crystals of acetylcholine receptor. J Cell Biol. 1984 Oct;99(4 Pt 1):1202–1211. doi: 10.1083/jcb.99.4.1202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brisson A., Wade R. H. Three-dimensional structure of luminal plasma membrane protein from urinary bladder. J Mol Biol. 1983 May 5;166(1):21–36. doi: 10.1016/s0022-2836(83)80048-5. [DOI] [PubMed] [Google Scholar]
  4. Buhle E. L., Jr, Knox B. E., Serpersu E., Aebi U. The structure of the Ca2+ ATPase as revealed by electron microscopy and image processing of ordered arrays. J Ultrastruct Res. 1983 Nov;85(2):186–203. doi: 10.1016/s0022-5320(83)90106-5. [DOI] [PubMed] [Google Scholar]
  5. Deisenhofer J., Epp O., Miki K., Huber R., Michel H. X-ray structure analysis of a membrane protein complex. Electron density map at 3 A resolution and a model of the chromophores of the photosynthetic reaction center from Rhodopseudomonas viridis. J Mol Biol. 1984 Dec 5;180(2):385–398. doi: 10.1016/s0022-2836(84)80011-x. [DOI] [PubMed] [Google Scholar]
  6. Dixon J. F., Hokin L. E. Studies on the characterization of the sodium-potassium transport adenosine triphosphatase. Purification and properties of the enzyme from the electric organ of Electrophorus electricus. Arch Biochem Biophys. 1974 Aug;163(2):749–758. doi: 10.1016/0003-9861(74)90537-2. [DOI] [PubMed] [Google Scholar]
  7. Engel A., Massalski A. 3D reconstruction from electron micrographs: its potential and practical limitations. Ultramicroscopy. 1984;13(1-2):71–83. doi: 10.1016/0304-3991(84)90058-5. [DOI] [PubMed] [Google Scholar]
  8. Fuller S. D., Capaldi R. A., Henderson R. Structure of cytochrome c oxidase in deoxycholate-drived two-dimensional crystals. J Mol Biol. 1979 Oct 25;134(2):305–327. doi: 10.1016/0022-2836(79)90037-8. [DOI] [PubMed] [Google Scholar]
  9. Hebert H., Jørgensen P. L., Skriver E., Maunsbach A. B. Crystallization patterns of membrane-bound (Na+ +K+)-ATPase. Biochim Biophys Acta. 1982 Aug 12;689(3):571–574. doi: 10.1016/0005-2736(82)90316-9. [DOI] [PubMed] [Google Scholar]
  10. Hebert H., Skriver E., Hegerl R., Maunsbach A. B. Structure of two-dimensional crystals of membrane-bound Na,K-ATPase as analyzed by correlation averaging. J Ultrastruct Res. 1985 Jul-Aug;92(1-2):28–35. doi: 10.1016/0889-1605(85)90124-7. [DOI] [PubMed] [Google Scholar]
  11. Herbert H., Skriver E., Maunsbach A. B. Three-dimensional structure of renal Na,K-ATPase determined by electron microscopy of membrane crystals. FEBS Lett. 1985 Jul 22;187(1):182–186. doi: 10.1016/0014-5793(85)81238-2. [DOI] [PubMed] [Google Scholar]
  12. Hokin L. E., Dahl J. L., Deupree J. D., Dioxon J. F., Hackney J. F., Perdue J. F. Studies on the characterization of the sodium-potassium transport adenosine triphosphatase. X. Purification of the enzyme from the rectal gland of Squalus acanthias. J Biol Chem. 1973 Apr 10;248(7):2593–2605. [PubMed] [Google Scholar]
  13. Jacob J. S., Miller K. R. Structure of a bacterial photosynthetic membrane. Isolation, polypeptide composition, and selective proteolysis. Arch Biochem Biophys. 1983 May;223(1):282–290. doi: 10.1016/0003-9861(83)90593-3. [DOI] [PubMed] [Google Scholar]
  14. Jorgensen P. L. Purification and characterization of (Na+ plus K+ )-ATPase. 3. Purification from the outer medulla of mammalian kidney after selective removal of membrane components by sodium dodecylsulphate. Biochim Biophys Acta. 1974 Jul 12;356(1):36–52. doi: 10.1016/0005-2736(74)90292-2. [DOI] [PubMed] [Google Scholar]
  15. Jørgensen P. L. Mechanism of the Na+, K+ pump. Protein structure and conformations of the pure (Na+ +K+)-ATPase. Biochim Biophys Acta. 1982 Aug 11;694(1):27–68. doi: 10.1016/0304-4157(82)90013-2. [DOI] [PubMed] [Google Scholar]
  16. Kawakami K., Noguchi S., Noda M., Takahashi H., Ohta T., Kawamura M., Nojima H., Nagano K., Hirose T., Inayama S. Primary structure of the alpha-subunit of Torpedo californica (Na+ + K+)ATPase deduced from cDNA sequence. Nature. 1985 Aug 22;316(6030):733–736. doi: 10.1038/316733a0. [DOI] [PubMed] [Google Scholar]
  17. Kawakami K., Nojima H., Ohta T., Nagano K. Molecular cloning and sequence analysis of human Na,K-ATPase beta-subunit. Nucleic Acids Res. 1986 Apr 11;14(7):2833–2844. doi: 10.1093/nar/14.7.2833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kyte J. Purification of the sodium- and potassium-dependent adenosine triphosphatase from canine renal medulla. J Biol Chem. 1971 Jul 10;246(13):4157–4165. [PubMed] [Google Scholar]
  19. Milligan R. A., Brisson A., Unwin P. N. Molecular structure determination of crystalline specimens in frozen aqueous solutions. Ultramicroscopy. 1984;13(1-2):1–9. doi: 10.1016/0304-3991(84)90051-2. [DOI] [PubMed] [Google Scholar]
  20. Mohraz M., Smith P. R. Structure of (Na+,K+)-ATPase as revealed by electron microscopy and image processing. J Cell Biol. 1984 May;98(5):1836–1841. doi: 10.1083/jcb.98.5.1836. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mohraz M., Yee M., Smith P. R. Novel crystalline sheets of Na,K-ATPase induced by phospholipase A2. J Ultrastruct Res. 1985 Oct-Nov;93(1-2):17–26. doi: 10.1016/0889-1605(85)90081-3. [DOI] [PubMed] [Google Scholar]
  22. Mohraz M., Yee M., Smith P. R. Structural studies of Na,K-ATPase. Ann N Y Acad Sci. 1986;483:131–139. doi: 10.1111/j.1749-6632.1986.tb34506.x. [DOI] [PubMed] [Google Scholar]
  23. Noguchi S., Noda M., Takahashi H., Kawakami K., Ohta T., Nagano K., Hirose T., Inayama S., Kawamura M., Numa S. Primary structure of the beta-subunit of Torpedo californica (Na+ + K+)-ATPase deduced from the cDNA sequence. FEBS Lett. 1986 Feb 17;196(2):315–320. doi: 10.1016/0014-5793(86)80270-8. [DOI] [PubMed] [Google Scholar]
  24. Ohta T., Nagano K., Yoshida M. The active site structure of Na+/K+-transporting ATPase: location of the 5'-(p-fluorosulfonyl)benzoyladenosine binding site and soluble peptides released by trypsin. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2071–2075. doi: 10.1073/pnas.83.7.2071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ovchinnikov YuA, Demin V. V., Barnakov A. N., Kuzin A. P., Lunev A. V., Modyanov N. N., Dzhandzhugazyan K. N. Three-dimensional structure of (Na+ + K+)-ATPase revealed by electron microscopy of two-dimensional crystals. FEBS Lett. 1985 Oct 7;190(1):73–76. doi: 10.1016/0014-5793(85)80430-0. [DOI] [PubMed] [Google Scholar]
  26. Robinson J. D., Flashner M. S. The (Na+ + K+)-activated ATPase. Enzymatic and transport properties. Biochim Biophys Acta. 1979 Aug 17;549(2):145–176. doi: 10.1016/0304-4173(79)90013-2. [DOI] [PubMed] [Google Scholar]
  27. Shull G. E., Lane L. K., Lingrel J. B. Amino-acid sequence of the beta-subunit of the (Na+ + K+)ATPase deduced from a cDNA. Nature. 1986 May 22;321(6068):429–431. doi: 10.1038/321429a0. [DOI] [PubMed] [Google Scholar]
  28. Shull G. E., Schwartz A., Lingrel J. B. Amino-acid sequence of the catalytic subunit of the (Na+ + K+)ATPase deduced from a complementary DNA. Nature. 1985 Aug 22;316(6030):691–695. doi: 10.1038/316691a0. [DOI] [PubMed] [Google Scholar]
  29. Skriver E., Maunsbach A. B., Jørgensen P. L. Formation of two-dimensional crystals in pure membrane-bound Na+,K+-ATPase. FEBS Lett. 1981 Aug 31;131(2):219–222. doi: 10.1016/0014-5793(81)80371-7. [DOI] [PubMed] [Google Scholar]
  30. Smith P. R., Fowler W. E., Pollard T. D., Aebi U. Structure of the actin molecule determined from electron micrographs of crystalline actin sheets with a tentative alignment of the molecule in the actin filament. J Mol Biol. 1983 Jul 5;167(3):641–660. doi: 10.1016/s0022-2836(83)80103-x. [DOI] [PubMed] [Google Scholar]
  31. Taylor K. A., Dux L., Martonosi A. Three-dimensional reconstruction of negatively stained crystals of the Ca2+-ATPase from muscle sarcoplasmic reticulum. J Mol Biol. 1986 Feb 5;187(3):417–427. doi: 10.1016/0022-2836(86)90442-0. [DOI] [PubMed] [Google Scholar]
  32. Unwin P. N., Ennis P. D. Two configurations of a channel-forming membrane protein. Nature. 1984 Feb 16;307(5952):609–613. doi: 10.1038/307609a0. [DOI] [PubMed] [Google Scholar]
  33. Williams R. C., Fisher H. W. Electron microscopy of tobacco mosaic virus under conditions of minimal beam exposure. J Mol Biol. 1970 Aug 28;52(1):121–123. doi: 10.1016/0022-2836(70)90181-6. [DOI] [PubMed] [Google Scholar]
  34. Zampighi G., Corless J. M., Robertson J. D. On gap junction structure. J Cell Biol. 1980 Jul;86(1):190–198. doi: 10.1083/jcb.86.1.190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zampighi G., Kyte J., Freytag W. Structural organization of (Na+ + K+)-ATPase in purified membranes. J Cell Biol. 1984 May;98(5):1851–1864. doi: 10.1083/jcb.98.5.1851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Zampighi G., Simon S. A., Kyte J., Kreman M. One-dimensional crystals of (Na+ + K+)-ATPase dimers. Biochim Biophys Acta. 1986 Jan 16;854(1):45–57. doi: 10.1016/0005-2736(86)90063-5. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES