Cytoplasmic amino acids within the membrane interface region influence connexin oligomerization

J Membr Biol. 2012 Jun;245(5-6):221-30. doi: 10.1007/s00232-012-9443-5. Epub 2012 Jun 22.

Abstract

Gap junction channels composed of connexins connect cells, allowing intercellular communication. Their cellular assembly involves a unique quality-control pathway. Some connexins [including connexin43 (Cx43) and Cx46] oligomerize in the trans-Golgi network following export of stabilized monomers from the endoplasmic reticulum (ER). In contrast, other connexins (e.g., Cx32) oligomerize early in the secretory pathway. Amino acids near the cytoplasmic aspect of the third transmembrane domain have previously been shown to determine this difference in assembly sites. Here, we characterized the oligomerization of two connexins expressed prominently in the vasculature, Cx37 and Cx40, using constructs containing a C-terminal dilysine-based ER retention/retrieval signal (HKKSL) or treatment with brefeldin A to block ER vesicle trafficking. Both methods led to intracellular retention of connexins, since the cells lacked gap junction plaques. Retention of Cx40 in the ER prevented it from oligomerizing, comparable to Cx43. By contrast, ER-retained Cx37 was partially oligomerized. Replacement of two amino acids near the third transmembrane domain of Cx43 (L152 and R153) with the corresponding amino acids from Cx37 (M152 and G153) resulted in early oligomerization in the ER. Thus, residues that allow Cx37 to oligomerize early in the secretory pathway could restrict its interactions with coexpressed Cx40 or Cx43 by favoring homomeric oligomerization, providing a structural basis for cells to produce gap junction channels with different connexin composition.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acids / chemistry
  • Amino Acids / metabolism*
  • Cell Membrane / metabolism*
  • Connexin 43 / chemistry
  • Connexin 43 / metabolism
  • Connexins / chemistry
  • Connexins / metabolism*
  • Cytosol / metabolism*
  • Fluorescent Antibody Technique
  • Gap Junction alpha-4 Protein
  • Gap Junctions / metabolism
  • Humans
  • Protein Multimerization
  • Structure-Activity Relationship

Substances

  • Amino Acids
  • Connexin 43
  • Connexins