pH regulation of connexin43: Molecular analysis of the gating particle

José F. Ek-Vitorín, Guillermo Calero, Gregory E. Morley, Wanda Coombs, Steven M. Taffet, Mario Delmar

Research output: Contribution to journalArticle

137 Scopus citations

Abstract

Gap junction channels allow for the passage of ions and small molecules between neighboring cells. These channels are formed by multimers of an integral membrane protein named connexin. In the heart and other tissues, the most abundant connexin is a 43-kDa, 382-amino acid protein termed connexin43 (Cx43). A characteristic property of connexin channels is that they close upon acidification of the intracellular space. Previous studies have shown that truncation of the carboxyl terminal of Cx43 impairs pH sensitivity. In the present study, we have used a combination of optical, electrophysiological, and molecular biological techniques and the oocyte expression system to further localize the regions of the carboxyl terminal that are involved in pH regulation of Cx43 channels. Our results show that regions 261-300 and 374-382 are essential components of a pH-dependent 'gating particle,' which is responsible for acidification-induced uncoupling of Cx43-expressing cells. Regions 261-300 and 374-382 seem to be interdependent. The function of region 261-300 may be related to the presence of a poly-proline repeat between amine acids 274 and 285. Furthermore, site- directed mutagenesis studies show that the function of region 374-382 is not directly related to its net balance of charges, although mutation of only one amine acid (aspartate 379) for asparagine impairs pH sensitivity to the same extent as truncation of the carboxyl terminal domain (from amine acid 257). The mutation in which serine 364 is substituted for proline, which has been associated with some cases of cardiac congenital malformations in humans, also disrupts the pH gating of Cx43, although deletion of amine acids 364- 373 has no effect on acidification-induced uncoupling. These results provide new insight into the molecular mechanisms responsible for acidification- induced uncoupling of gap junction channels in the heart and in other Cx43- expressing structures.

Original languageEnglish (US)
Pages (from-to)1273-1284
Number of pages12
JournalBiophysical Journal
Volume71
Issue number3
DOIs
StatePublished - Sep 1996
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics

Fingerprint Dive into the research topics of 'pH regulation of connexin43: Molecular analysis of the gating particle'. Together they form a unique fingerprint.

  • Cite this