Abstract
Basic principles of the gating mechanisms of neuronal sodium channels, especially the fast inactivation process, were revealed by a quantitative analysis of the effects of the chemically irreversible modifying agent chloramine T. The compound is known to enhance the open probability of sodium channels by interfering with the inactivation process. The key for the deduction of structure–function relationships was obtained from the analysis of single-channel patch-clamp data, especially the finding that chloramine T-induced modification of inactivation occurred in four steps. These steps were termed modes 1–4 (four-mode gating model), and their temporal sequence was always the same. The kinetic analysis of single-channel traces with an improved two-dimensional dwell-time fit revealed the possible mechanism related to each mode. Similarities to the kinetics of the sodium channel mutant F1489Q led to the assignment of modes 1 and 2 to transient defects in the locking of the inactivation particle (hinged lid). In the third mode, the hinged lid was unable to lock permanently. Finally, in mode 4, the apparent single-channel current was reduced, which could be explained by fast gating, presumably related to the selectivity filter.
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Abbreviations
- 2D fit:
-
two-dimensional dwell-time fit
- 2D histogram:
-
two-dimensional dwell-time histogram
- 4M-model:
-
four-mode gating model
- CT:
-
chloramine T
- SNR:
-
signal-to-noise ratio
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Acknowledgment
We are grateful to S. Schnell and D. Gremelle for technical assistance. We thank W. A. Catterall and T. Scheuer for Nav1.2a and Nav1.2a F1489Q cDNA and W. Ulbricht for helpful discussions. This work was supported by the Bundesministerium für Bildung und Forschung (03F0261A) and the Deutsche Forschungsgemeinschaft (Ha 712/11-3 and Ha712/14-2).
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Huth, T., Schmidtmayer, J., Alzheimer, C. et al. Four-mode gating model of fast inactivation of sodium channel Nav1.2a. Pflugers Arch - Eur J Physiol 457, 103–119 (2008). https://doi.org/10.1007/s00424-008-0500-y
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DOI: https://doi.org/10.1007/s00424-008-0500-y