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Aminoglycoside block of P2X2 receptors heterologously expressed in Xenopus laevis oocytes

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Abstract

Aminoglycosides are polycationic antibiotics that have been shown to block a variety of cation channels. The inhibitory effect of externally applied aminoglycosides on P2X2 receptor currents was examined after heterologous expression in Xenopus laevis oocytes using the two-electrode voltage-clamp technique. All of the aminoglycosides tested inhibited the ATP-evoked responses with potencies ranging from 71 μM to 2 mM (IC50 values). The ranked order of potency was streptomycin > gentamicin > neomycin > paromomycin > kanamycin. The inhibition of P2X receptor currents was independent of the ATP concentration used for the activation, which is compatible with a noncompetitive mechanism. The inhibition was voltage-dependent and was reduced at more positive membrane potentials. To examine whether the current block was dependent on the receptor conformation, the aminoglycoside effect on a non-desensitizing P2X2-X1 receptor chimera was analyzed. The results from these measurements suggest that inhibition is caused by an open pore block that locks the P2X receptor chimera in an open nonconducting state from which the agonist dissociation is slow. We also demonstrate that the P2X2-X1 chimera can serve as a tool to directly test whether an antagonist acts competitively or not.

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Acknowledgements

We thank Anja Becker and Janna Enderich for technical assistance, Ernst Bamberg for generous support, and Günther Schmalzing for critical review of the manuscript.

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Authors and Affiliations

  1. Department of Biophysical Chemistry, Max Planck Institute of Biophysics, Max-von-Laue-Strasse 3, 60438, Frankfurt, Germany

    Eva-Verena Bongartz & Jürgen Rettinger

  2. Molecular Pharmacology, RWTH Aachen University, Wendlingweg 2, 52074, Aachen, Germany

    Ralf Hausmann

Authors
  1. Eva-Verena Bongartz
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  2. Jürgen Rettinger
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  3. Ralf Hausmann
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Correspondence to Ralf Hausmann.

Additional information

Jürgen Rettinger and Ralf Hausmann contributed equally to this work.

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Bongartz, EV., Rettinger, J. & Hausmann, R. Aminoglycoside block of P2X2 receptors heterologously expressed in Xenopus laevis oocytes. Purinergic Signalling 6, 393–403 (2010). https://doi.org/10.1007/s11302-010-9204-9

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