Selectivity of connexin 43 channels is regulated through protein kinase C-dependent phosphorylation.
Coordinated contractile activation of the heart and resistance to ischemic injury depend, in part, on the intercellular communication mediated by Cx43-composed gap junctions. The function of these junctions is regulated at multiple levels (assembly to degradation) through phosphorylation at specific sites in the carboxyl terminus (CT) of the Cx43 protein. We show here that the selective permeability of Cx43 junctions is regulated through protein kinase C (PKC)-dependent phosphorylation at serine 368 (S368). Selective permeability was measured in several Cx43-expressing cell lines as the rate constant for intercellular dye diffusion relative to junctional conductance. The selective permeability of Cx43 junctions under control conditions was quite variable, as was the open-state behavior of the comprising channels. Coexpression of the CT of Cx43 as a distinct protein, treatment with a PKC inhibitor, or mutation of S368 to alanine, all reduced (or eliminated) phosphorylation at S368, reduced the incidence of 55- to 70-pS channels, and reduced by 10-fold the selective permeability of the junctions for a small cationic dye. Because PKC activation during preischemic conditioning is cardioprotective during subsequent ischemic episodes, we examined no-flow, ischemic hearts for Cx43 phosphorylated at S368 (pS368). Consistent with early activation of PKC, pS368-Cx43 was increased in ischemic hearts; despite extensive lateralization of total Cx43, pS368-Cx43 remained predominantly at intercalated disks. Our data suggest that the selectivity of gap junction channels at intercalated disks is increased early in ischemia.
Ek-Vitorin
Jose F
JF
Department of Physiology, University of Arizona, Tucson 85724, USA.
King
Timothy J
TJ
Heyman
Nathanael S
NS
Lampe
Paul D
PD
Burt
Janis M
JM
D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D004396:Coloring Agents / Q000493:pharmacokinetics; D018031:Connexin 43 / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D017629:Gap Junctions / Q000378:metabolism*; D007365:Intercellular Junctions / Q000378:metabolism; D007473:Ion Channels / Q000378:metabolism*; D051379:Mice; D008815:Mice, Inbred Strains; D017202:Myocardial Ischemia / Q000378:metabolism*; D010539:Permeability; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats
D004396:Coloring Agents; D018031:Connexin 43; D007473:Ion Channels; D011493:Protein Kinase C