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Inactivation of L-type calcium channels is determined by the length of the N terminus of mutant β1 subunits

  • Ion Channels, Receptors and Transporters
  • Published:
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Abstract

Voltage-dependent calcium channel (Cav) pores are modulated by cytosolic β subunits. Four β-subunit genes and their splice variants offer a wide structural array for tissue- or disease-specific biophysical gating phenotypes. For instance, the length of the N terminus of β2 subunits has major effects on activation and inactivation rates. We tested whether a similar mechanism principally operates in a β1 subunit. Wild-type β1a subunit (N terminus length 60 aa) and its newly generated N-terminal deletion mutants (51, 27 and 18 aa) were examined within recombinant L-type calcium channel complexes (Cav1.2 and α2δ2) in HEK293 cells at the whole-cell and single-channel level. Whole-cell currents were enhanced by co-transfection of the full-length β1a subunit and by all truncated constructs. Voltage dependence of steady-state activation and inactivation did not depend on N terminus length, but inactivation rate was diminished by N terminus truncation. This was confirmed at the single-channel level, using ensemble average currents. Additionally, gating properties were estimated by Markov modeling. In confirmation of the descriptive analysis, inactivation rate, but none of the other transition rates, was reduced by shortening of the β1a subunit N terminus. Our study shows that the length-dependent mechanism of modulating inactivation kinetics of β2 calcium channel subunits can be confirmed and extended to the β1 calcium channel subunit.

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Acknowledgements

The authors gratefully acknowledge Dr. Dirk Gründemann for fruitful discussion to construct β1a-subunit mutants, Dr. Gyula Varadi for generously providing the HEK293α1c cell line, Katja Witschas and Peter Bartels for their excellent support of electrophysiological measurements, Ramin Akhavan-Malyer for a preliminary Markov modeling approach, and Dr. Alexey Kuzmenkin for his large contribution to the IonFit software development. E.K. is indebted to the Christiane Nuesslein-Volhard Foundation for the financial support. This work was supported by the Center of Molecular Medicine Cologne (CMMC A4 to S.H.).

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Authors and Affiliations

  1. Department of Pharmacology, University of Cologne, Gleueler Strasse 24, 50931, Cologne, Germany

    Wanchana Jangsangthong, Elza Kuzmenkina, Ismail F. Y. Khan, Jan Matthes & Stefan Herzig

  2. Center for Molecular Medicine, University of Cologne, Robert-Koch-Str. 21, 50931, Cologne, Germany

    Wanchana Jangsangthong & Stefan Herzig

  3. Centre for cardiovascular and metabolic diseases, University of Lausanne, Rue du Bugnon 21, 1005, Lausanne, Switzerland

    Roger Hullin

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  1. Wanchana Jangsangthong
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  2. Elza Kuzmenkina
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  3. Ismail F. Y. Khan
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  4. Jan Matthes
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  5. Roger Hullin
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  6. Stefan Herzig
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Correspondence to Stefan Herzig.

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Jangsangthong, W., Kuzmenkina, E., Khan, I.F.Y. et al. Inactivation of L-type calcium channels is determined by the length of the N terminus of mutant β1 subunits. Pflugers Arch - Eur J Physiol 459, 399–411 (2010). https://doi.org/10.1007/s00424-009-0738-z

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  • DOI: https://doi.org/10.1007/s00424-009-0738-z

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