Novel crystalline sheets of Na,K-ATPase induced by phospholipase A2

Mohraz, Manijeh; Yee, Movien; Smith, Paul

doi:10.1016/0889-1605(85)90081-3

Cited by 30 publications

(17 citation statements)

References 27 publications

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“…Significant changes in the fluorescence of lucifer-labeled (Na,K)-ATPase were observed after the addition of ligand combinations known to induce crystallization of (Na,K)-ATPase in the plane of the membrane (Skriver et al, 1981;Zampighi et al, 1984; Mohraz et al, 1985). Figure 4 shows two representative traces of the time course of the fluorescence changes after addition of Mn, Mg, and Pi to lucifer-labeled the @-subunit of another a@ dimer is unlikely to have been significant because the distance of closest approach between two a@ dimers is about twice the Ro value of 29 A (Moczydlowski & Fortes, 1981c;Hebert et al, 1985;Ovchinnikov et al, 1985).…”

Section: Discussionmentioning

confidence: 97%

Spatial relationship and conformational changes between the cardiac glycoside site and .beta.-subunit oligosaccharides in sodium plus potassium activated adenosinetriphosphatase

Lee¹,

Fortes²

1986

Biochemistry

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(Na,K)-ATPase, the enzyme responsible for active transport of Na and K across the plasma membranes of animal cells, consists of a catalytic subunit (alpha) and a glycoprotein subunit (beta) with unknown function. We have determined the distance between fluorescent probes directed to specific sites on the alpha- and beta-subunits and ligand-induced changes in the fluorescence of a probe specifically attached to the beta-subunit. The cardiac glycoside site on the alpha-subunit was labeled with anthroylouabain [Fortes, P. A. G. (1977) Biochemistry 16, 531-540]. The oligosaccharides on the beta-subunit were labeled with lucifer yellow carbohydrazide [Lee, J. A., & Fortes, P. A. G. (1985) Biochemistry 24, 322-330]. Resonance energy transfer from anthroylouabain to lucifer yellow was measured by steady-state and time-resolved fluorescence spectroscopy. The distance between these probes was determined from the efficiency of energy transfer. The average distance between anthroylouabain and lucifer yellow was 47 A and was independent of the number of acceptor molecules attached to the beta-subunit. The measured distance corresponds to the distance between the cardiac glycoside site and the center of the labeled oligosaccharides on the beta-subunit within one alpha beta dimer. The distance was the same (47 A) when anthroylouabain was bound with ATP or Pi as phosphorylating ligands but increased to 49 A in the presence of vanadate. The change in average distance provides quantitative evidence of a conformational difference between the complexes of cardiac glycosides with (Na,K)-ATPase induced by phosphorylating ligands or by vanadate.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Discussionmentioning

confidence: 97%

Spatial relationship and conformational changes between the cardiac glycoside site and .beta.-subunit oligosaccharides in sodium plus potassium activated adenosinetriphosphatase

Lee¹,

Fortes²

1986

Biochemistry

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“…The size of the scanned area is 300 X 300 nm 2 , and the dimensions of the unit cell of the two dimensional crystals were 8 X 10 nm 2 , with an angle of 75°between the rows. These structures are interpreted as dimers of the Na,K-ATPase in accordance with Mohraz et al (1985). The buckling of the fragment in the upper right corner of the image has been caused by underlying debris adsorbed to the mica.…”

Section: Comparison Of Preparationsmentioning

confidence: 99%

“…Crystallization of the protein is possible in both the E. and E 2 conformations and as monomers or dimers in the unit cell, depending on the composition of the crystallization buffer (Maunsbach et al, 1991). Besides vanadate or phosphate, phospholipase A 2 has been used to induce the crystallization (Mohraz et al, 1985). It has been shown that crystallization occurs as the result of hydrolysis and solubilization of the phospholipids of the membrane fragments.…”

Section: Crystallizationmentioning

confidence: 99%

Investigation of the Na,K-ATPase by SFM

Apell

Colchero²,

Linder³

et al. 1993

STM and SFM in Biology

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“…The unit cell parameters may vary significantly even within the same specimen batch. Modification of the membranes with phospholipase AZ results in dimeric crystals [4]. Incubation with an ATP-analogue, Co(NH&ATP, which stabilizes a cobalt-tetraminephosphoenzyme, gives rise to tetragonal p4 crystals [5].…”

Section: Introductionmentioning

confidence: 99%

Coexistence of different forms ofNa,K‐ATPase in two‐dimensional membrane crystals

et al. 1990

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Two-dimensional membrane crystals of renal Na,K-ATPase were analyzed by electron microscopy and image processing. The particular property of the crystals in this work was that they showed unit cell parameters similar to the previously studied p21 crystals but lacked the dyad axis as observed in nominal O"-projections. A three-dimensional reconstruction revealed that structural differences between ag-units of the enzyme gave rise to the asymmetry. A high degree of two-fold rotational symmetry was observed in the middle of the structure while the protein units had different three-dimensional shapes at levels above and below the central sections. The simultaneous coexistence of different forms of Na,K-ATPase suggests that the conformational flexibility of the enzyme plays an important role in the pumping process.

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Novel crystalline sheets of Na,K-ATPase induced by phospholipase A2

Cited by 30 publications

References 27 publications

Spatial relationship and conformational changes between the cardiac glycoside site and .beta.-subunit oligosaccharides in sodium plus potassium activated adenosinetriphosphatase

Spatial relationship and conformational changes between the cardiac glycoside site and .beta.-subunit oligosaccharides in sodium plus potassium activated adenosinetriphosphatase

Investigation of the Na,K-ATPase by SFM

Coexistence of different forms ofNa,K‐ATPase in two‐dimensional membrane crystals

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