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Biochemistry (Moscow)

, Volume 81, Issue 3, pp 249–254 | Cite as

Identification of a region of the polypeptide chain of Na,K-ATPase α-subunit interacting with 67-kDa melittin-like protein

  • Yu. V. Kamanina
  • E. A. Klimanova
  • E. A. Dergousova
  • I. Yu. Petrushanko
  • O. D. LopinaEmail author
Article
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Abstract

It was shown earlier that a 67-kDa protein purified from mouse kidney using polyclonal antibodies against melittin (a peptide from bee venom) interacted with Na,K-ATPase from rabbit kidney. In this study, a 43-kDa proteolytic fragment of Na,K-ATPase α-subunit interacting with the 67-kDa melittin-like protein was found. The α-subunit was hydrolyzed by trypsin in the presence of 0.5 mM ouabain (E2-conformation of Na,K-ATPase). A proteolytic fragment interacting with the 67-kDa melittin-like protein that was identified by mass-spectrometry is a region of the cytoplasmic domain of Na,K-ATPase α-subunit located between amino acid residues 591 and 775. The fragment includes a conservative DPPRA motif that occurs in many P-type ATPases. It was shown earlier that this motif of H,K-ATPase from gastric mucosa binds to melittin. We suggest that namely this motif of P-type ATPases is able to interact with proteins containing melittin-like modules.

Key words

Na,K-ATPase melittin-like proteins protein—protein interactions 

Abbreviations

CHAPS

3-[(3-cholamidoprolyl)dimethylammonium]-1-propanesulfonate

ELISA

enzyme-linked immunosorbent assay

PMSF

phenylmethylsulfonyl fluoride

PBS

phosphate-buffered saline

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References

  1. 1.
    Lopina, O. D. (2000) Na+,K+-ATPase: structure, mechanism, and regulation, Membr. Cell Biol., 13, 721–744.PubMedGoogle Scholar
  2. 2.
    Geering, K., Beggah, A., Good, P., Girardet, S., Roy, S., Schaer, D., and Jaunin, P. (1996) Oligomerization and maturation of Na,K-ATPase: functional interaction of the cytoplasmic NH2 terminus of the β subunit with the α-subunit, J. Cell Biol., 133, 1193–1204.CrossRefPubMedGoogle Scholar
  3. 3.
    Arystarkhova, E., Wetzel, R. K., Asinovski, N. K., and Sweadner, K. J. (1999) The γ-subunit modulates Na+ and K+ affinity of the renal Na,K-ATPase, J. Biol. Chem., 274, 33183–33185.CrossRefPubMedGoogle Scholar
  4. 4.
    Halsey, J. F., Mountcastle, D. B., Takeguchi, C. A., Biltonen, R. L., and Lindenmayer, G. E. (1977) Detection of a ouabain-induced structural change in the sodium, potassium-adenosine triphosphatase, Biochemistry, 16, 432–435.CrossRefPubMedGoogle Scholar
  5. 5.
    Koob, R., Kraemer, D., Trippe, G., Aebi, U., and Drenckhahn, D. (1990) Association of kidney and parotid Na+,K+-ATPase microsomes with actin and analogs of spectrin, and ankyrin, Eur. J. Cell Biol., 53, 93–100.PubMedGoogle Scholar
  6. 6.
    Yudowski, G. A., Efendiev, R., Pedemonte, R., Katz, A. I., Berggren, P. O., and Bertorello, A. M. (2000) Phosphoinositide-3 kinase binds to a proline-rich motif in the Na+,K+-ATPase α-subunit and regulates its trafficking, Proc. Natl. Acad. Sci. USA, 97, 6556–6561.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Ferrandi, M., Salardi, S., Tripodi, G., Barassi, P., Rivera, R., Manunta, P., Goldshleger, R., Ferrari, P., Bianchi, G., and Karlish, S. J. (1999) Evidence for an interaction between adducin and Na+-K+-ATPase: relation to genetic hypertension, Am. J. Physiol., 277, 1338–1349.Google Scholar
  8. 8.
    Liu, J., Kesiry, R., Periyasamy, S. M., Malhotra, D., Xie, Z., and Shapiro, J. I. (2004) Ouabain induces endocytosis of plasmalemmal Na/K-ATPase in LLC-PK1 cells by a clathrin-dependent mechanism, Kidney Int., 66, 227–241.CrossRefPubMedGoogle Scholar
  9. 9.
    Wang, H., Haas, M., Liang, M., Cai, T., Tian, J., Li, S., and Xie, Z. (2004) Ouabain assembles signaling cascades through the caveolar Na+/K+-ATPase, J. Biol. Chem., 279, 17250–17259.CrossRefPubMedGoogle Scholar
  10. 10.
    Lee, K., Jung, J., Kim, M., and Guidotti, G. (2001) Interaction of the α-subunit of Na,K-ATPase with cofilin, Biochem. J., 353, 377–385.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Dolgova, N. V., Kamanina, Y. V., Akimova, O. A., Orlov, S. N., Rubtsov, A. M., and Lopina, O. D. (2007) A protein whose binding to Na,K-ATPase is regulated by ouabain, Biochemistry (Moscow), 72, 863–871.CrossRefGoogle Scholar
  12. 12.
    Kaetzel, M. A., and Dedman, J. R. (1987) Identification of a 55-kDa high-affinity calmodulin-binding protein from Electrophorus electricus, J. Biol. Chem., 262, 3726–3729.PubMedGoogle Scholar
  13. 13.
    Cuppoletti, J., and Abbot, A. J. (1990) Interaction of melittin with the Na+,K+-ATPase: evidence for a melittin-induced conformational change, Arch. Biochem. Biophys., 283, 249–257.CrossRefPubMedGoogle Scholar
  14. 14.
    Cuppoletti, J. (1990) [125I]Azidosalicylyl-melittin-binding domains: evidence for a polypeptide receptor on the gastric (H++K+)ATPase, Arch. Biochem. Biophys., 278, 405–415.CrossRefGoogle Scholar
  15. 15.
    Jorgensen, P. L., and Skou, J. C. (1971) Purification and characterization of (Na++K+)-ATPase. I. The influence of detergents on the activity of (Na++K+)-ATPase in preparations from the outer medulla of rabbit kidney, Biochim. Biophys. Acta, 233, 366–380.CrossRefPubMedGoogle Scholar
  16. 16.
    Schagger, H., and Von Jagow, G. (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa, Anal. Biochem., 166, 368–379.CrossRefPubMedGoogle Scholar
  17. 17.
    Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature, 259, 680–685.CrossRefGoogle Scholar
  18. 18.
    Jorgensen, P. L. (1977) Purification and characterization of (Na++K+)-ATPase. VI. Differential tryptic modification of catalytic functions of the purified enzyme in presence of NaCl and KCl, Biochim. Biophys. Acta, 466, 97–108.CrossRefPubMedGoogle Scholar
  19. 19.
    Karlish, S. J. (1980) Characterization of conformational changes in Na,K-ATPase labeled with fluorescein at the active site, J. Bioenerg. Biomembr., 12, 111–136.CrossRefPubMedGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2016

Authors and Affiliations

  • Yu. V. Kamanina
    • 1
    • 2
  • E. A. Klimanova
    • 1
    • 2
  • E. A. Dergousova
    • 1
    • 2
  • I. Yu. Petrushanko
    • 2
  • O. D. Lopina
    • 1
    Email author
  1. 1.Department of Biochemistry, School of BiologyLomonosov Moscow State UniversityMoscowRussia
  2. 2.Engelhardt Institute of Molecular BiologyRussian Academy of SciencesMoscowRussia

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