The permanently charged lidocaine analogue QX222 acts as a blocker from the intracellular side and...
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MEETING ABSTRACT Open Access The permanently charged lidocaine analogue QX222 acts as a blocker from the intracellular side and as an inactivation modulator from the extracellular side in a mutant Na V 1.4 channel Péter Lukács, René Cervenka, Xaver Koenig, Agnés Mike, Touran Zarrabi, Karlheinz Hilber, Hannes Todt * From 16th Scientific Symposium of the Austrian Pharmacological Society (APHAR) Vienna, Austria. 25-27 November 2010 Background QX222 is a quaternary amine analogue of lidocaine, which, unlike lidocaine, is permanently charged. Lido- caine has its binding site in the internal vestibule of the voltage-gated sodium channel. Due to the hydrophobic nature of its uncharged form, lidocaine reaches the binding site by passing through the membrane, QX222 can reach this binding site only by a hydrophilic path- way, presumably through the channel protein. However, such a pathway has been reported only in the heart-type sodium channel (Na V 1.5) and some mutants of other sodium channels. Notably, mutations at site 1575 in the skeletal muscle-type sodium channel (Na V 1.4) open an access pathway from the external side. In this study we tested the properties of QX222 block on the mutant I1575E. Methods All measurements were done in tsA201 cells transiently transfected with the Na V 1.4 sodium channel a subunit, cotransfected with b1 sodium channel subunit. Currents were recorded by patch-clamp technique in whole-cell configuration. Results Both 500 μM lidocaine and 500 μM QX222 shifted the half-point of steady-state slow inactivation to hyperpo- larized potentials in I1575E if applied from the extracel- lular side. However, only lidocaine significantly shifted the half-point of steady-state fast inactivation. Intracellu- lar application of QX222 resulted in a quick block of sodium current, indicating that the drug entered the channel, but the hyperpolarizing shift of steady-state fast inactivation was still not present. In addition, with intra- cellular application of QX222, the strong hyperpolariz- ing shift in steady-state slow inactivation disappeared. Conclusions These results suggest that the binding site for use- dependent block is in the inner vestibule of the channel, that fast inactivation is modulated only by the hydro- phobic form of local anaesthetics, and that the binding site for modulation of slow inactivation is only accessi- ble form the extracellular side of the channel. Acknowledgements This work was supported by grants P21006-B11, P17509-B11, P13961-B05 from the FWF. Published: 16 November 2010 doi:10.1186/1471-2210-10-S1-A24 Cite this article as: Lukács et al.: The permanently charged lidocaine analogue QX222 acts as a blocker from the intracellular side and as an inactivation modulator from the extracellular side in a mutant Na V 1.4 channel. BMC Pharmacology 2010 10(Suppl 1):A24. * Correspondence: [email protected] Institute of Pharmacology, Center of Physiology and Pharmacology, Medical University of Vienna, 1090 Vienna, Austria Lukács et al. BMC Pharmacology 2010, 10(Suppl 1):A24 http://www.biomedcentral.com/1471-2210/10/S1/A24 © 2010 Todt et al; licensee BioMed Central Ltd.
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Transcript of The permanently charged lidocaine analogue QX222 acts as a blocker from the intracellular side and...
MEETING ABSTRACT Open Access
The permanently charged lidocaine analogueQX222 acts as a blocker from the intracellularside and as an inactivation modulator from theextracellular side in a mutant NaV1.4 channelPéter Lukács, René Cervenka, Xaver Koenig, Agnés Mike, Touran Zarrabi, Karlheinz Hilber, Hannes Todt*
From 16th Scientific Symposium of the Austrian Pharmacological Society (APHAR)Vienna, Austria. 25-27 November 2010
BackgroundQX222 is a quaternary amine analogue of lidocaine,which, unlike lidocaine, is permanently charged. Lido-caine has its binding site in the internal vestibule of thevoltage-gated sodium channel. Due to the hydrophobicnature of its uncharged form, lidocaine reaches thebinding site by passing through the membrane, QX222can reach this binding site only by a hydrophilic path-way, presumably through the channel protein. However,such a pathway has been reported only in the heart-typesodium channel (NaV1.5) and some mutants of othersodium channels. Notably, mutations at site 1575 in theskeletal muscle-type sodium channel (NaV1.4) open anaccess pathway from the external side. In this study wetested the properties of QX222 block on the mutantI1575E.
MethodsAll measurements were done in tsA201 cells transientlytransfected with the NaV1.4 sodium channel a subunit,cotransfected with b1 sodium channel subunit. Currentswere recorded by patch-clamp technique in whole-cellconfiguration.
ResultsBoth 500 μM lidocaine and 500 μM QX222 shifted thehalf-point of steady-state slow inactivation to hyperpo-larized potentials in I1575E if applied from the extracel-lular side. However, only lidocaine significantly shifted
the half-point of steady-state fast inactivation. Intracellu-lar application of QX222 resulted in a quick block ofsodium current, indicating that the drug entered thechannel, but the hyperpolarizing shift of steady-state fastinactivation was still not present. In addition, with intra-cellular application of QX222, the strong hyperpolariz-ing shift in steady-state slow inactivation disappeared.
ConclusionsThese results suggest that the binding site for use-dependent block is in the inner vestibule of the channel,that fast inactivation is modulated only by the hydro-phobic form of local anaesthetics, and that the bindingsite for modulation of slow inactivation is only accessi-ble form the extracellular side of the channel.
AcknowledgementsThis work was supported by grants P21006-B11, P17509-B11, P13961-B05from the FWF.
Published: 16 November 2010
doi:10.1186/1471-2210-10-S1-A24Cite this article as: Lukács et al.: The permanently charged lidocaineanalogue QX222 acts as a blocker from the intracellular side and as aninactivation modulator from the extracellular side in a mutant NaV1.4channel. BMC Pharmacology 2010 10(Suppl 1):A24.
* Correspondence: [email protected] of Pharmacology, Center of Physiology and Pharmacology, MedicalUniversity of Vienna, 1090 Vienna, Austria
Lukács et al. BMC Pharmacology 2010, 10(Suppl 1):A24http://www.biomedcentral.com/1471-2210/10/S1/A24
© 2010 Todt et al; licensee BioMed Central Ltd.
