J Membr Biol 2000 Nov 01;1781:55-70. doi: 10.1007/s002320010015.

Slow gating of gap junction channels and calmodulin.

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Certain COOH-terminus mutants of connexin32 (Cx32) were previously shown to form channels with unusual transjuctional voltage (V(j)) sensitivity when tested heterotypically in oocytes against Cx32 wild type. Junctional conductance (G(j)) slowly increased by severalfold or decreases to nearly zero with V(j) positive or negative, respectively, at mutant side, and V(j) positive at mutant side reversed CO(2)-induced uncoupling. This suggested that the CO(2)-sensitive gate might be a V(j)-sensitive slow gate. Based on previous data for calmodulin (CaM) involvement in gap junction function, we have hypothesized that the slow gate could be a CaM-like pore plugging molecule (cork gating model). This study describes a similar behavior in heterotypic channels between Cx32 and each of four new Cx32 mutants modified in cytoplasmic-loop and/or COOH-terminus residues. The mutants are: ML/NN+3R/N, 3R/N, ML/NN and ML/EE; in these mutants, N or E replace M105 and L106, and N replace R215, R219 and R220. This study also reports that inhibition of CaM expression strongly reduces V(j) and CO(2) sensitivities of two of the most effective mutants, suggesting a CaM role in slow and chemical gating.

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Species referenced: Xenopus laevis
Genes referenced: gjb1