Conformational maturation and post-er multisubunit assembly of gap junction proteins

Judy K. VanSlyke, Christian C. Naus, Linda Musil

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

For all previously well-characterized oligomeric integral membrane proteins, folding, multisubunit assembly, and recognition of conformationally immature molecules for degradation occurs at their organelle of synthesis. This cannot, however, be the case for the gap junction-forming protein connexin43 (Cx43), which when endogenously expressed undergoes multisubunit assembly into connexons only after its transport to the trans-Golgi network. We have developed two novel assays to assess Cx43 folding and assembly: acquisition of resistance of disulfide bonds to reduction by extracellularly added DTT and Triton X-114 detergent phase partitioning. We show that Cx43 synthesized at physiologically relevant levels undergoes a multistep conformational maturation process in which folding of connexin monomers within the ER is a prerequisite for multisubunit assembly in the TGN. Similar results were obtained with Cx32, disproving the widely reported contention that the site of endogenous -connexin assembly is the ER. Exogenous overexpression of Cx43, Cx32, or Cx26 allows these events to take place within the ER, the first example of the TGN and ER as alternative sites for oligomeric assembly. Our findings also constitute the first biochemical evidence that defective connexin folding is a cause of the human disorder X-linked Charcot-Marie-Tooth disease.

Original languageEnglish (US)
Pages (from-to)2451-2463
Number of pages13
JournalMolecular Biology of the Cell
Volume20
Issue number9
DOIs
StatePublished - May 1 2009

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Connexin 43
Connexins
trans-Golgi Network
Charcot-Marie-Tooth Disease
Protein Folding
Disulfides
Organelles
Detergents
Membrane Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Conformational maturation and post-er multisubunit assembly of gap junction proteins. / VanSlyke, Judy K.; Naus, Christian C.; Musil, Linda.

In: Molecular Biology of the Cell, Vol. 20, No. 9, 01.05.2009, p. 2451-2463.

Research output: Contribution to journalArticle

VanSlyke, Judy K. ; Naus, Christian C. ; Musil, Linda. / Conformational maturation and post-er multisubunit assembly of gap junction proteins. In: Molecular Biology of the Cell. 2009 ; Vol. 20, No. 9. pp. 2451-2463.
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