Role of histidine 95 on pH gating of the cardiac gap junction protein connexin43

Jose F. Ek, Mario Delmar, Raisa Perzova, Steven M. Taffet

Research output: Contribution to journalArticlepeer-review

81 Scopus citations


We have studied the role of histidine 95 (H95) on the pH gating of the cardiac gap junction protein connexin43 (Cx43). Wild-type and mutant rat cardiac Cx43 channels were expressed in antisense-injected Xenopus oocytes. Junctional conductance was measured using the dual voltage-clamp technique, and intracellular acidification was induced by superfusion with a sodium acetate-containing solution balanced at a pH of 6.2. H95 was substituted by other amino acids by use of oligonucleotide-directed site-specific mutagenesis. Replacing H95 for the hydrophobic residues methionine or phenylalanine, for the charged basic residue arginine, or for the noncharged residue glutamine (H95Q) yielded nonfunctional channels. Functional expression of H95Q was rescued by placing a histidine residue in position 93 (H95Q-L93H), 94 (H95Q-A94H), or 97 (H95Q-F97H) but not in position 96. Further experiments showed that replacing H95 with either aspartate (an acidic residue) or tyrosine (a polar uncharged residue) led to the expression of functional channels with a reduced susceptibility to acidification- induced uncoupling, whereas lysine (a basic residue) was more susceptible to uncoupling than the wild-type protein. The susceptibility to acidification- induced uncoupling was enhanced for the H95Q-A94H mutant when compared with the wild-type mutant, but it was significantly reduced when histidine was placed at position 93 (H95Q-L93H). Our data indicate that a property placed histidine residue is an important structural element for functional expression as well as for pH regulation of Cx43. The results suggest that the importance of H95 on pH gating may be associated with a possible protonation of this residue on acidification of the intracellular environment. The data are compatible with a recently proposed mechanism modeled after the 'ball- and-chain' hypothesis-explaining pH gating of Cx43.

Original languageEnglish (US)
Pages (from-to)1058-1064
Number of pages7
JournalCirculation research
Issue number6
StatePublished - Jun 1994


  • cardiac gap junctions
  • connexin43
  • histidine
  • pH(i)
  • site-directed mutagenesis

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine


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