Three-dimensional structure of Na,K-ATPase determined from membrane crystals induced by cobalt-tetrammine-ATP

Skriver, Elisabeth; Kavéus, Urban; Hebert, Hans; Maunsbach, Arvid B.

doi:10.1016/1047-8477(92)90017-5

Cited by 18 publications

(5 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…On the cytoplasmic side of the membrane, the Na ϩ ,K ϩ -ATPase ␣ subunit consists of a stalk just above the membrane, which is connected to a tripartite head extending ϳ75 Å above the membrane. Generally speaking, this structure is consistent with previous reconstructions of Na ϩ ,K ϩ -ATPase (Mohraz et al, 1987;Skriver et al, 1992;Hebert et al, 1985Hebert et al, , 1988, but with more clearly defined domains and membrane topology. The cytoplasmic headpiece is reminiscent of that from Ca 2ϩ -ATPase in the E 2 conformation, which consists of a highly conserved phosphorylation domain (P) sitting on top of a narrow stalk with a nose pointing to one side and a nucleotide-binding domain (N) above (Toyoshima et al, 2000).…”

Section: Overall Shape Of Na ؉ K ؉ -Atpasesupporting

confidence: 90%

“…Previous comparisons of 8-Å structures of Neurospora H ϩ -ATPase and rabbit muscle Ca 2ϩ -ATPase from cryoelectron microscopy (Kühlbrandt et al, 1998; provided a similar view of the conformational change, though specific assignment of the cytoplasmic domains was ambiguous at that time. With respect to Na ϩ ,K ϩ -ATPase, although it was the first member of this family to be discovered (Skou, 1957) and the first to form twodimensional (2D) crystals (Skriver et al, 1981), structural models have been limited to ϳ25-Å resolution (Mohraz et al, 1987;Skriver et al, 1992;Hebert et al, 1985Hebert et al, , 1988, because of poor order, small crystal size, and the use of negative stain. Here we present a structure of E 2 -state Na ϩ ,K ϩ -ATPase from cryoelectron microscopy at 11-Å resolution, offering the first view of the entire heterodimer with unambiguous assignment of its domains.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Structure of Na+,K+-ATPase at 11-Å Resolution: Comparison withCa2+-ATPase in E1 and E2 States

Rice

Young

Martin

et al. 2001

Biophysical Journal

View full text Add to dashboard Cite

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.

show abstract

Section: Overall Shape Of Na ؉ K ؉ -Atpasesupporting

confidence: 90%

Section: Introductionmentioning

confidence: 99%

Structure of Na+,K+-ATPase at 11-Å Resolution: Comparison withCa2+-ATPase in E1 and E2 States

Rice

Young

Martin

et al. 2001

Biophysical Journal

View full text Add to dashboard Cite

show abstract

“…These and the present findings are also similar to those of an FTIR study on the proteolytically cleaved nicotinic acetylcholine receptor (5), from which it was concluded that the transmembrane domains were relatively rich in ␤-structures, in agreement with suggestions based on electron microscope image reconstructions (27). Currently, electron crystallography of the Na,K-ATPase does not yet approach this sort of resolution (28). In the present study, it is not known to what extent the relatively high content of ␤-type structures of the trypsinized enzyme may be attributable to a rearrangement of the secondary structure in the parts of the tryptic fragments that remain external to the membrane.…”

Section: Discussionsupporting

confidence: 79%

Characterization of the Secondary Structure and Assembly of the Transmembrane Domains of Trypsinized Na,K-ATPase by Fourier Transform Infrared Spectroscopy

Heimburg¹,

Esmann²,

Marsh³

1997

Journal of Biological Chemistry

View full text Add to dashboard Cite

Fourier transform infrared spectroscopy has been used to compare native Na,K-ATPase-containing membranes with those trypsinized in the presence of either Rb ؉ or Na ؉ ions to remove the extramembranous parts of the protein. The protein secondary structure content deduced from the amide I band is approximately 30 -35% ␣-helix, 37-40% ␤-structure, and 13-15% random coil for native membranes from shark rectal gland and from pig kidney, in both the Na-and K-forms. Trypsinization in either Rb ؉ (a K ؉ congener) or Na ؉ removes approximately 35% of the amide I band intensity of native membranes from shark rectal gland. The protein secondary structural content of the trypsinized membranes lies in the range of approximately 23-32% ␣-helix, 37-46% ␤-structure, and 12-18% random coil for the shark and kidney enzymes. The distribution of intensity between the bands corresponding to protonated and deuteriumexchanged ␣-helices, and between the component bands attributed to ␤-structure, changes considerably on trypsinization, in the direction of a greater proportion of protonated ␣-helix and a broader range of frequencies for ␤-structure. The kinetics of deuteration of the slowly exchanging population of protein amide groups is also changed on trypsinization. The mean rate constant for deuteration of trypsinized membranes is approximately half that for native membranes, whereas the proportion of amides contributing to this population increases on trypsinization. The temperature dependence of the amide I band in the Fourier transform infrared spectra indicates that the onset of thermal denaturation occurs at 58°C for native membranes (in either Na ؉ or K ؉ ) and for membranes trypsinized in Rb ؉ , but the major denaturation event for membranes trypsinized in Na ؉ occurs at approximately 84°C. These results correlate with the functional properties of the intramembranous section of the enzyme.

show abstract

“…However, it was later discovered that decavanadate was actually responsible for Ca 2+ -ATPase crystallization (48,60,96) and that the same Na + /K + -ATPase crystal form could also be induced by phospholipase A2 in the absence of vanadate (63). A variety of other two-dimensional crystals were studied, including those from scallop adductor muscle in both E 2 (11) and E 2 -P (35) conformations and from Na + /K + -ATPase stabilized by ATP analogs (86). In addition, both Ca 2+ -ATPase (25) and H + -ATPase (17) were crystallized in the E 1 conformation.…”

Section: Electron Microscopy Of Ca 2+ -Atpase and Other P-type Atpasesmentioning

confidence: 99%

Structure and Function of the Calcium Pump

Stokes

Green

2003

Annu. Rev. Biophys. Biomol. Struct.

View full text Add to dashboard Cite

Active transport of cations is achieved by a large family of ATP-dependent ion pumps, known as P-type ATPases. Various members of this family have been targets of structural and functional investigations for over four decades. Recently, atomic structures have been determined for Ca2+-ATPase by X-ray crystallography, which not only reveal the architecture of these molecules but also offer the opportunity to understand the structural mechanisms by which the energy of ATP is coupled to calcium transport across the membrane. This energy coupling is accomplished by large-scale conformational changes. The transmembrane domain undergoes plastic deformations under the influence of calcium binding at the transport site. Cytoplasmic domains undergo dramatic rigid-body movements that deliver substrates to the catalytic site and that establish new domain interfaces. By comparing various structures and correlating functional data, we can now begin to associate the chemical changes constituting the reaction cycle with structural changes in these domains.

show abstract

Three-dimensional structure of Na,K-ATPase determined from membrane crystals induced by cobalt-tetrammine-ATP

Cited by 18 publications

References 29 publications

Structure of Na+,K+-ATPase at 11-Å Resolution: Comparison withCa2+-ATPase in E1 and E2 States

Structure of Na+,K+-ATPase at 11-Å Resolution: Comparison withCa2+-ATPase in E1 and E2 States

Characterization of the Secondary Structure and Assembly of the Transmembrane Domains of Trypsinized Na,K-ATPase by Fourier Transform Infrared Spectroscopy

Structure and Function of the Calcium Pump

Contact Info

Product

Resources

About