Stimulation, Inhibition, or Stabilization of Na,K-ATPase Caused by Specific Lipid Interactions at Distinct Sites

Habeck, Michael; Haviv, Haim; Katz, Adriana; Kapri-Pardes, Einat; Ayciriex, Sophie; Shevchenko, Andrej; Ogawa, Hirotaka; Toyoshima, Chikashi; Karlish, Steven J. D.

doi:10.1074/jbc.m114.611384

Cited by 59 publications

(92 citation statements)

References 70 publications

(90 reference statements)

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“…It is the molecular mechanism that remains elusive. As we have reported recently, the activity of recombinant Na, K-ATPase prepared with sphingomyelin and cholesterol is inhibited by 70 -80%, due to a strong reduction in the molar turnover rate (20). Taken together, the findings of the Na,KATPase⅐caveolin complex and inhibition by sphingomyelin/ cholesterol could be compatible with the presence of heterogeneous populations of the pump in the cell membrane.…”

Section: Possible Mechanisms Of Nak-atpase-mediated Cell Signaling?supporting

confidence: 73%

Do Src Kinase and Caveolin Interact Directly with Na,K-ATPase?

Yosef¹,

Katz²,

Peleg³

et al. 2016

Journal of Biological Chemistry

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Much evidence points to a role of Na,K-ATPase in ouabaindependent signal transduction. Based on experiments with different cell lines and native tissue membranes, a current hypothesis postulates direct interactions between the Na,K-ATPase and Src kinase (non-receptor tyrosine kinase). Na,K-ATPase is proposed to bind Src kinase and inhibit its activity, whereas ouabain, the specific Na,K-ATPase inhibitor, binds and stabilizes the E2 conformation, thus exposing the Src kinase domain and its active site Tyr-418 for activation. Ouabain-dependent signaling is thought to be mediated within caveolae by a complex consisting of Na,K-ATPase, caveolin, and Src kinase. In the current work, we have looked for direct interactions utilizing purified recombinant Na,K-ATPase (human ␣1␤1FXYD1 or porcine ␣1D369N␤1FXYD1) and purified human Src kinase and human caveolin 1 or interactions between these proteins in native membrane vesicles isolated from rabbit kidney. By several independent criteria and techniques, no stable interactions were detected between Na,K-ATPase and purified Src kinase. Na,KATPase was found to be a substrate for Src kinase phosphorylation at Tyr-144. Clear evidence for a direct interaction between purified human Na,K-ATPase and human caveolin was obtained, albeit with a low molar stoichiometry (1:15-30 caveolin 1/Na,K-ATPase). In native renal membranes, a specific caveolin 1 4 -5 oligomer (95 kDa) was found to be in direct interaction with Na,K-ATPase. We inferred that a small fraction of the renal Na,K-ATPase molecules is in a ϳ1:1 complex with a caveolin 1 4 -5 oligomer. Thus, overall, whereas a direct caveolin 1/Na,K-ATPase interaction is confirmed, the lack of direct Src kinase/Na,K-ATPase binding requires reassessment of the mechanism of ouabain-dependent signaling.

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Section: Possible Mechanisms Of Nak-atpase-mediated Cell Signaling?supporting

confidence: 73%

Do Src Kinase and Caveolin Interact Directly with Na,K-ATPase?

Yosef¹,

Katz²,

Peleg³

et al. 2016

Journal of Biological Chemistry

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“…In this study, we did not observe significant stimulation or inhibition of the basal ATPase activity by lipids as shown for the Na,KATPase (5,25). From a molecular point of view, this could suggest preferential interactions between McjD and negatively charged lipids.…”

Section: Discussionmentioning

confidence: 53%

Structural and Functional Basis for Lipid Synergy on the Activity of the Antibacterial Peptide ABC Transporter McjD

Mehmood

Corradi

Choudhury

et al. 2016

Journal of Biological Chemistry

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“…Stopped-Flow Experiments with RH421-Stopped-flow measurements (27) were performed with an Applied Photophysics SX20 system. To detect electrogenic reaction steps with RH421, the monochromator for excitation was set to 577 Ϯ 6 nm.…”

Section: Methodsmentioning

confidence: 99%

“…Recombinant Human Na,K-ATPase Expression and Purification-P. pastoris transformation, yeast growth, membrane preparation, and His tag purification of recombinant human ␣1␤1FXYD1 and ␣2␤1FXYD1 Na,K-ATPase were done essentially as described previously (27)(28)(29)(30). Purified detergent-soluble Na,K-ATPase complexes were eluted from BD-Talon beads in a medium containing 250 mM imidazole, pH 7.2, 10% glycerol, 100 mM NaCl, 25 mM Tris-Tricine, pH 7.4, 0.12 mg/ml C 12 E 8 , 0.025 mg/ml cholesterol, 0.075 mg/ml SOPS, and 0.075 mg/ml SOPC.…”

Section: Methodsmentioning

confidence: 99%