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Phosphorylation in the C-terminus of the rat connexin46 (rCx46) and regulation of the conducting activity of the formed connexons

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Abstract

To analyse the role of PKC-dependent phosphorylation in the C-terminus of rCx46 in regulation of rCx46 connexons, truncated mutants rCx4645.3 and rCx4644.2 which end before and after PKC-dependent phosphorylation sites respectively were generated. Both rCx4645.3 and rCx4644.2 formed connexons in Xenopus oocytes similar to Cx46wt-connexons. They were activated by depolarisation above −40 mV and at voltages above 50 mV, inactivation was spontaneously observed or induced by PKC activator TPA, suggesting that inactivation does not require PKC-dependent phosphorylation in the C-terminus. Three casein-kinase-II-(CKII)-dependent phosphorylation sites were also identified. rCx4637.7 and rCx4628.2 respectively without two or all of these sites were generated. rCx4637.7-connexons were similar to rCx46wt-connexons. rCx4628.2-connexons comparable to rCx46wt-connexons were observed after injection of 50 times more rCx4628.2-mRNA (25 ng per oocyte). CKII-blocker inhibited depolarisation-evoked currents in oocytes injected with 0.5 ng per oocyte rCx4637.7-mRNA or rCx46wt-mRNA. Injection of 25 ng per oocyte rCx4637.7-mRNA or rCx46wt-mRNA overcame the effect of CKII-inhibitor. We propose that CKII-dependent phosphorylation in the C-terminus accelerates formation of rCx46-connexons.

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Authors and Affiliations

  1. Institute of Biophysics, Leibniz University Hannover, Herrenhäuserstr. 2, 30419, Hannover, Germany

    Carsten Zeilinger, Willem Bintig, Hans-Albert Kolb & Anaclet Ngezahayo

  2. Molecular and Cell Physiology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    Wilhelm J. Walter

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  1. Wilhelm J. Walter
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  2. Carsten Zeilinger
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  3. Willem Bintig
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  4. Hans-Albert Kolb
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  5. Anaclet Ngezahayo
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Correspondence to Anaclet Ngezahayo.

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Walter, W.J., Zeilinger, C., Bintig, W. et al. Phosphorylation in the C-terminus of the rat connexin46 (rCx46) and regulation of the conducting activity of the formed connexons. J Bioenerg Biomembr 40, 397–405 (2008). https://doi.org/10.1007/s10863-008-9151-0

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  • DOI: https://doi.org/10.1007/s10863-008-9151-0

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