Mutational analysis of gap junction formation

Gerhard Dahl, R. Werner, E. Levine, C. Rabadan-Diehl

Research output: Contribution to journalArticle

125 Citations (Scopus)

Abstract

The paired oocyte cell-cell channel assay was used to investigate the mechanisms involved in the process of formation of gap junction channels. Single oocytes, injected with connexin-specific mRNAs, accumulate a pool of precursors from which cell-cell channels can form rapidly upon pairing. Several lines of evidence, including immunohistochemistry and surface labeling, indicate that part of this precursor pool is located in the cell membrane, probably in the form of closed hemichannels. The homophilic binding of hemichannels to each other can be mimicked by synthetic peptides representing the extracellular loop sequences of connexin32. The peptides specifically suppress channel formation. A crucial role is established for the six cysteines in the extracellular domains that are conserved in all vertebrate gap junction proteins. Change of any of these cysteines into serines results in absolute loss of function of the mutant connexin. The effects of thiol-specific reagents on channel formation suggest that docking and/or opening of channels involves disulfide exchange. Several of the variable amino acids in the extracellular loop sequences were found to determine specificity of connexin-connexin interactions.

Original languageEnglish
Pages (from-to)172-182
Number of pages11
JournalBiophysical Journal
Volume62
Issue number1
StatePublished - Jan 1 1992

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Connexins
Gap Junctions
Oocytes
Cysteine
Sulfhydryl Reagents
Peptides
Disulfides
Serine
Vertebrates
Immunohistochemistry
Cell Membrane
Amino Acids
Messenger RNA

ASJC Scopus subject areas

  • Biophysics

Cite this

Dahl, G., Werner, R., Levine, E., & Rabadan-Diehl, C. (1992). Mutational analysis of gap junction formation. Biophysical Journal, 62(1), 172-182.

Mutational analysis of gap junction formation. / Dahl, Gerhard; Werner, R.; Levine, E.; Rabadan-Diehl, C.

In: Biophysical Journal, Vol. 62, No. 1, 01.01.1992, p. 172-182.

Research output: Contribution to journalArticle

Dahl, G, Werner, R, Levine, E & Rabadan-Diehl, C 1992, 'Mutational analysis of gap junction formation', Biophysical Journal, vol. 62, no. 1, pp. 172-182.
Dahl G, Werner R, Levine E, Rabadan-Diehl C. Mutational analysis of gap junction formation. Biophysical Journal. 1992 Jan 1;62(1):172-182.
Dahl, Gerhard ; Werner, R. ; Levine, E. ; Rabadan-Diehl, C. / Mutational analysis of gap junction formation. In: Biophysical Journal. 1992 ; Vol. 62, No. 1. pp. 172-182.
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