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Palytoxin Acts on Na+,K+-ATPase but not Nongastric H+,K+-ATPase

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Abstract

Palytoxin (PTX) opens a pathway for ions to pass through Na,K-ATPase. We investigate here whether PTX also acts on nongastric H,K-ATPases. The following combinations of cRNA were expressed in Xenopus laevis oocytes: Bufo marinus bladder H,K-ATPase α2- and Na,K-ATPase β2-subunits; Bufo Na,K-ATPase α1- and Na,K-ATPase β2-subunits; and Bufo Na,K-ATPase β2-subunit alone. The response to PTX was measured after blocking endogenous Xenopus Na,K-ATPase with 10 μm ouabain. Functional expression was confirmed by measuring 86Rb uptake. PTX (5 nm) produced a large increase of membrane conductance in oocytes expressing Bufo Na,K-ATPase, but no significant increase occurred in oocytes expressing Bufo H,K-ATPase or in those injected with Bufo β2-subunit alone. Expression of the following combinations of cDNA was investigated in HeLa cells: rat colonic H,K-ATPase α1-subunit and Na,K-ATPase β1-subunit; rat Na,K-ATPase α2-subunit and Na,K-ATPase β2-subunit; and rat Na,K-ATPase β1- or Na,K-ATPase β2-subunit alone. Measurement of increases in 86Rb uptake confirmed that both rat Na,K and H,K pumps were functional in HeLa cells expressing rat colonic HKα1/NKβ1 and NKα2/NKβ2. Whole-cell patch-clamp measurements in HeLa cells expressing rat colonic HKα1/NKβ1 exposed to 100 nm PTX showed no significant increase of membrane current, and there was no membrane conductance increase in HeLa cells transfected with rat NKβ1- or rat NKβ2-subunit alone. However, in HeLa cells expressing rat NKα2/NKβ2, outward current was observed after pump activation by 20 mm K+ and a large membrane conductance increase occurred after 100 nm PTX. We conclude that nongastric H,K-ATPases are not sensitive to PTX when expressed in these cells, whereas PTX does act on Na,K-ATPase.

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References

  • Artigas P, Gadsby DC (2003) Na+/K+-pump ligands modulate gating of palytoxin-induced ion channels. Proc Natl Acad Sci USA 100:501–505

    Article  PubMed  CAS  Google Scholar 

  • Artigas P, Gadsby DC (2004) Large diameter of palytoxin-induced Na/K pump channels and modulation of palytoxin interaction by Na/K pump ligands. J Gen Physiol 123:357–376

    Article  PubMed  CAS  Google Scholar 

  • Burnay M, Crambert G, Geering K, Horisberger J-D (2001) Bufo marinus bladder H,K-ATPase carries out electroneutral ion transport. Am J Physiol 281:F869–F874

    CAS  Google Scholar 

  • Caplan MJ (1997) Ion pumps in epithelial cells: sorting, stabilization, and polarity. Am J Physiol 272:G1304–G1313

    PubMed  CAS  Google Scholar 

  • Cantley LG, Resh MD, Guidoti G (1978) Vanadate inhibits the red cell Na,K-ATPase from the cytoplasmic side. Nature 272:552–554

    Article  PubMed  CAS  Google Scholar 

  • Codina J, Kone BC, Delmas-Mata JT, Dubose TD Jr (1996) Functional expression of the H+,K+-ATPase α subunit. J Biol Chem 271:29759–29763

    Article  PubMed  CAS  Google Scholar 

  • Cougnon M, Planelles G, Crowson MS, Shull GE, Rossier B, Jaisser F (1996) The rat distal colon P-ATPase α-subunit encodes a ouabain-sensitive H,K-ATPase. J Biol Chem 271:7277–7280

    Article  PubMed  CAS  Google Scholar 

  • Cougnon M, Bouyer P, Planelles G, Jaisser F (1998) Does the colonic H,K-ATPase also act as an Na,K-ATPase? Proc Natl Acad Sci USA 95:6516–6520

    Article  PubMed  CAS  Google Scholar 

  • Crowson MS, Shull GE (1992) Isolation and characterization of a cDNA encoding the putative distal colon H+,K+-ATPase. J Biol Chem 267:13740–13748

    PubMed  CAS  Google Scholar 

  • Del Castillo JR, Rajendran VM, Binder HJ (1991) Apical membrane localization of ouabain-sensitive K+-activated ATPase activities in rat distal colon. Am J Physiol 261:G1005–G1011

    PubMed  Google Scholar 

  • Geering K (1998) Molecular mechanisms involved in the regulation of Na,K-ATPase expression. Adv Mol Cell Biol 23B:275–309

    CAS  Google Scholar 

  • Gottardi CJ, Caplan MJ (1993) Delivery of Na,K-ATPase in polarized epithelial cells. Science 260:552–554

    Article  PubMed  CAS  Google Scholar 

  • Grishen AV, Sverdlov VE, Costina MB, Modyanov NN (1994) Cloning and characterization of the entire cDNA encoded by ATP1AL1–a member of the human Na,K/H,K-ATPase gene family. FEBS Lett 349:144–150

    Article  Google Scholar 

  • Guennoun S, Horisberger J-D (2000) Structure of the 5th transmembrane segment of the Na,K-ATPase alpha subunit: a cysteine-scanning mutagenesis study. FEBS Lett 482:144–148

    Article  PubMed  CAS  Google Scholar 

  • Guennoun S, Horisberger J-D (2002) Cysteine-scanning mutagenesis study of the sixth transmembrane segment of the Na,K-ATPase alpha subunit. FEBS Lett 513:277–281

    Article  PubMed  CAS  Google Scholar 

  • Guennoun S, Rakowski RF, Horisberger J-D (2005) Bufo marinus H,K-ATPase is not sensitive to palytoxin. J Gen Physiol 126:58a

    Google Scholar 

  • Habermann E (1989) Palytoxin acts through Na+,K+ ATPase. Toxicon 27:1171–1187

    Article  PubMed  CAS  Google Scholar 

  • Hilgemann DW (2003) From a pump to a pore: how palytoxin opens the gates. Proc Natl Acad Sci USA 100:386–388

    Article  PubMed  CAS  Google Scholar 

  • Horisberger J-D, Kharoubi-Hess S, Guennoun S, Michielin O (2004) The fourth transmembrane segment of the Na,K-ATPase α subunit. J Biol Chem 279:29542–29550

    Article  PubMed  CAS  Google Scholar 

  • Marti-Renom MA, Stuart AC, Fiser A, Sanchez R, Melo F, Sali A (2000) Comparative protein structure modeling of genes and genomes. Annu Rev Biophys Biomol Struct 29:291–325

    Article  PubMed  CAS  Google Scholar 

  • Mullin JM, Snock KV, McGinn MT (1991) Effects of apical vs. basolateral palytoxin on LLC-PK1 renal epithelia. Am J Physiol 260:C1201–C1211

    PubMed  CAS  Google Scholar 

  • Munson KR, Garcia G, Sachs G (2005) Inhibitor and ion binding sites on the gastric H,K-ATPase. Biochemistry 44:5267–5284

    Article  PubMed  CAS  Google Scholar 

  • Ozaki H, Nagase H, Urakawa N (1985) Interaction of palytoxin and cardiac glycosides on erythrocytes membrane and (Na+,K+)ATPase. Eur J Biochem 152:475–480

    Article  PubMed  CAS  Google Scholar 

  • Pestov NB, Korneenko TV, Adams G, Tillekeratne M, Shakhparonov MI, Modyanov NN (2002) Nongastric H-K-ATPase in rodent prostate: lobe-specific expression and apical localization. Am J Physiol 282:C907–C916

    CAS  Google Scholar 

  • Post RL, Jolly PC (1957) The linkage of sodium, potassium, and ammonium active transport across the human erythrocyte membrane. Biochim Biophys Acta 25:118–128

    Article  PubMed  CAS  Google Scholar 

  • Rabon EC, McFall TL, Sachs G (1982) The gastric [H,K]ATPase: H+/ATP stoichiometry. J Biol Chem 257:6296–6299

    PubMed  CAS  Google Scholar 

  • Rakowski RF, Gadsby DC, De Weer P (1989) Stoichiometry and voltage dependence of the sodium pump in voltage-clamped, internally dialyzed squid giant axon. J Gen Physiol 93:903–941

    Article  PubMed  CAS  Google Scholar 

  • Rakowski RF, Paxson CL (1988) Voltage dependence of Na/K pump current in Xenopus oocytes. J Membr Biol 106:173–182

    Article  PubMed  CAS  Google Scholar 

  • Rakowski RF, Sagar S (2003) Found: Na and K binding sites of the sodium pump. News Physiol Sci 18:164–168

    PubMed  CAS  Google Scholar 

  • Sachs G, Chang HH, Rabon E, Schackman R, Lewin M, Saccomani G (1976) A nonelectrogenic H+ pump in plasma membranes of hog stomach. J Biol Chem 251:7690–7698

    PubMed  CAS  Google Scholar 

  • Sangan P, Thevananther S, Sangan S, Rajendran VM, Binder HJ (2000) Colonic H,K-ATPase alpha- and beta-subunits express ouabain-insensitive H,K-ATPase. Am J Physiol 278:C182–C189

    CAS  Google Scholar 

  • Scheiner-Bobis G (1998) Ion-transporting ATPases as ion channels. Naunyn Schmiedebergs Arch Pharmacol 357:477–482

    Article  PubMed  CAS  Google Scholar 

  • Scheiner-Bobis G, Hubschle T, Diener M (2002) Action of palytoxin on apical H+/K+-ATPase in rat colon. Eur J Biochem 269:3905–3911

    Article  PubMed  CAS  Google Scholar 

  • Silver RB, Soleimani M (1999) H+,K+-ATPases: regulation and role in pathophysiological states. Am J Physiol 276:F799–F811

    PubMed  CAS  Google Scholar 

  • Smolka M, Helander HF, Sachs G (1983) Monoclonal antibodies against gastric H+/K+ ATPase. Am J Physiol 245:G589–G596

    PubMed  CAS  Google Scholar 

  • Spicer Z, Clarke LL, Gawenis LR, Shull GE (2001) Colonic H+-K+-ATPase in K+ conservation and electrogenic Na+ absorption during Na+ restriction. Am J Physiol 281:G1369–G1377

    CAS  Google Scholar 

  • Sweadner KJ, Donnet C (2001) Structural similarities of Na,K-ATPase and SERCA, the Ca2+-ATPase of the sarcoplasmic reticulum. Biochem J 356:685–704

    Article  PubMed  CAS  Google Scholar 

  • Toyoshima C, Numora H, Tsuda T (2004) Lumenal gating mechanism revealed in calcium pump crystal structures with phosphate analogues. Nature 432:361–368

    Article  PubMed  CAS  Google Scholar 

  • Wang X, Horisberger J-D (1997) Palytoxin effects through interaction with the Na,K-ATPase in Xenopus oocyte. FEBS Lett 409:391–395

    Article  PubMed  CAS  Google Scholar 

  • Wang S, Takeyasu K (1997) Primary structure and evolution of the ATP-binding domains of the P-type ATPases in Tetrahymena thermophila. Am J Physiol 272:C715–C728

    PubMed  CAS  Google Scholar 

  • Wu CH, Vasilets LA, Takeda K, Kawamura M, Schwarz W (2003) Functional role of the N-terminus of Na+,K+-ATPase α-subunit as an inactivation gate of palytoxin-induced pump channel. Biochim Biophys Acta 1609:55–62

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

We thank Gary Shull for providing the rat colonic H,K-ATPase α-subunit and the Na,K-ATPase α1-subunit cDNAs. We also thank Robert Levenson for providing the rat Na,K-ATPase β1- and β2-subunit cDNAs. We also thank Joshua Berlin for assistance with the mammalian expression system and for providing facilities to start this project. We thank Kaethi Geering for providing suggestions for this work. This investigation was supported in part by funds from an American Heart Association postdoctoral fellowship (to S. G.-L.), National Institutes of Health grant NS-22979, National Science Foundation grant CCF-0622158 and Swiss National Fund grant 31-65441.01.

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Authors and Affiliations

  1. Department of Biological Sciences, Ohio University, Athens, OH, 45701, USA

    Saida Guennoun-Lehmann & Robert F. Rakowski

  2. School of Electrical Engineering and Computer Science, Ohio University, Athens, OH, 45701, USA

    James E. Fonseca

  3. Department of Pharmacology and Toxicology, University of Lausanne, Rue du Bugnon 27, CH-1005, Lausanne, Switzerland

    Jean-Daniel Horisberger

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  1. Saida Guennoun-Lehmann
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  2. James E. Fonseca
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  3. Jean-Daniel Horisberger
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  4. Robert F. Rakowski
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Correspondence to Robert F. Rakowski.

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Guennoun-Lehmann, S., Fonseca, J.E., Horisberger, JD. et al. Palytoxin Acts on Na+,K+-ATPase but not Nongastric H+,K+-ATPase. J Membrane Biol 216, 107–116 (2007). https://doi.org/10.1007/s00232-007-9040-1

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  • DOI: https://doi.org/10.1007/s00232-007-9040-1

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