Details
| Original language | English |
|---|---|
| Pages (from-to) | 397-405 |
| Number of pages | 9 |
| Journal | Journal of Bioenergetics and Biomembranes |
| Volume | 40 |
| Issue number | 4 |
| Publication status | Published - Aug 2008 |
Abstract
To analyse the role of PKC-dependent phosphorylation in the C-terminus of rCx46 in regulation of rCx46 connexons, truncated mutants rCx46(45.3) and rCx46(44.2) which end before and after PKC-dependent phosphorylation sites respectively were generated. Both rCx46(45.3) and rCx46(44.2) formed connexons in Xenopus oocytes similar to Cx46(wt)-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. rCx46(37.7) and rCx46(28.2) respectively without two or all of these sites were generated. rCx46(37.7)-connexons were similar to rCx46(wt)-connexons. rCx46(28.2)-connexons comparable to rCx46(wt)-connexons were observed after injection of 50 times more rCx46(28.2)-mRNA (25 ng per oocyte). CKII-blocker inhibited depolarisation-evoked currents in oocytes injected with 0.5 ng per oocyte rCx46(37.7)-mRNA or rCx46(wt)-mRNA. Injection of 25 ng per oocyte rCx46(37.7)-mRNA or rCx46(wt)-mRNA overcame the effect of CKII-inhibitor. We propose that CKII-dependent phosphorylation in the C-terminus accelerates formation of rCx46-connexons.
Keywords
- Animals, Cells, Cultured, Connexins/physiology, Gap Junctions/physiology, Ion Channel Gating/physiology, Membrane Potentials/physiology, Oocytes/physiology, Phosphorylation, Rats, Xenopus laevis
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In: Journal of Bioenergetics and Biomembranes, Vol. 40, No. 4, 08.2008, p. 397-405.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Phosphorylation in the C-terminus of the rat connexin46 (rCx46) and regulation of the conducting activity of the formed connexons
AU - Walter, Wilhelm J
AU - Zeilinger, Carsten
AU - Bintig, Willem
AU - Kolb, Hans-Albert
AU - Ngezahayo, Anaclet
N1 - Funding information: The work was partly supported by the project NANOTOME; Biophotonik III. Dr. Zeilinger was supported by the Helmholtzgemeinschaft Virtual Institute for Biological Structure Research (VIBS); VH-VI-013.
PY - 2008/8
Y1 - 2008/8
N2 - To analyse the role of PKC-dependent phosphorylation in the C-terminus of rCx46 in regulation of rCx46 connexons, truncated mutants rCx46(45.3) and rCx46(44.2) which end before and after PKC-dependent phosphorylation sites respectively were generated. Both rCx46(45.3) and rCx46(44.2) formed connexons in Xenopus oocytes similar to Cx46(wt)-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. rCx46(37.7) and rCx46(28.2) respectively without two or all of these sites were generated. rCx46(37.7)-connexons were similar to rCx46(wt)-connexons. rCx46(28.2)-connexons comparable to rCx46(wt)-connexons were observed after injection of 50 times more rCx46(28.2)-mRNA (25 ng per oocyte). CKII-blocker inhibited depolarisation-evoked currents in oocytes injected with 0.5 ng per oocyte rCx46(37.7)-mRNA or rCx46(wt)-mRNA. Injection of 25 ng per oocyte rCx46(37.7)-mRNA or rCx46(wt)-mRNA overcame the effect of CKII-inhibitor. We propose that CKII-dependent phosphorylation in the C-terminus accelerates formation of rCx46-connexons.
AB - To analyse the role of PKC-dependent phosphorylation in the C-terminus of rCx46 in regulation of rCx46 connexons, truncated mutants rCx46(45.3) and rCx46(44.2) which end before and after PKC-dependent phosphorylation sites respectively were generated. Both rCx46(45.3) and rCx46(44.2) formed connexons in Xenopus oocytes similar to Cx46(wt)-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. rCx46(37.7) and rCx46(28.2) respectively without two or all of these sites were generated. rCx46(37.7)-connexons were similar to rCx46(wt)-connexons. rCx46(28.2)-connexons comparable to rCx46(wt)-connexons were observed after injection of 50 times more rCx46(28.2)-mRNA (25 ng per oocyte). CKII-blocker inhibited depolarisation-evoked currents in oocytes injected with 0.5 ng per oocyte rCx46(37.7)-mRNA or rCx46(wt)-mRNA. Injection of 25 ng per oocyte rCx46(37.7)-mRNA or rCx46(wt)-mRNA overcame the effect of CKII-inhibitor. We propose that CKII-dependent phosphorylation in the C-terminus accelerates formation of rCx46-connexons.
KW - Animals
KW - Cells, Cultured
KW - Connexins/physiology
KW - Gap Junctions/physiology
KW - Ion Channel Gating/physiology
KW - Membrane Potentials/physiology
KW - Oocytes/physiology
KW - Phosphorylation
KW - Rats
KW - Xenopus laevis
U2 - 10.1007/s10863-008-9151-0
DO - 10.1007/s10863-008-9151-0
M3 - Article
C2 - 18668357
VL - 40
SP - 397
EP - 405
JO - Journal of Bioenergetics and Biomembranes
JF - Journal of Bioenergetics and Biomembranes
SN - 0145-479X
IS - 4
ER -