Analysis of ectatomin action on cell membranes

Kirill Pluzhnikov, Elena Nosyreva, Ludmila Shevchenko, Yuri Kokoz, Dirk Schmalz, Ferdinand Hucho, Evgeniy Grishin

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Abstract

Ectatomin (m = 7928 Da) is a toxic component from the Ectatomma tuberculatum ant venom containing two homologous polypeptide chains (37 and 34 residues) linked to each other by a disulfide bond. In aqueous solution it forms a four α-helix bundle. At concentrations of 0.05-0.1 μM, ectatomin forms channels in cellular and artificial bilayer membranes. Immunochemical analysis of the intracellular distribution of ectatomin showed that the toxin gets efficiently inserted into the plasma membrane at a concentration of 5 x 10-7 M and does not penetrate inside the cell. The effect of ectatomin on cardiac L-type calcium current was studied. Calcium currents (I(Ca)) in isolated rat cardiac ventricular myocytes were measured using the whole-cell perforated patch-clamp technique. It was shown that ectatomin at concentrations of 0.01-10 nM inhibited I(Ca) after a latency of few seconds. I(Ca) was decreased twofold by 10 nM ectatomin. However, the most prominent effect of ectatomin was observed after stimulation of I(Ca) by isoproterenol, an agonist of β-adrenoreceptors, or forskolin, a stimulator of adenylate cyclase. At a concentration of 1 nm, ectatomin abolished the isoproterenol- and forskolin-sensitive components of I(Ca). The inhibitory effect of ectatomin was partially reversed by subsequent application of 2 μM of forskolin, whereas subsequent isoproterenol application did not produce the same effect.

Original languageEnglish (US)
Pages (from-to)501-506
Number of pages6
JournalEuropean Journal of Biochemistry
Volume262
Issue number2
DOIs
Publication statusPublished - Jun 1 1999

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Keywords

  • Calcium channels
  • Cytotoxicity
  • Ectatomin
  • β-adrenergic receptor

ASJC Scopus subject areas

  • Biochemistry

Cite this

Pluzhnikov, K., Nosyreva, E., Shevchenko, L., Kokoz, Y., Schmalz, D., Hucho, F., & Grishin, E. (1999). Analysis of ectatomin action on cell membranes. European Journal of Biochemistry, 262(2), 501-506. https://doi.org/10.1046/j.1432-1327.1999.00426.x