Association of connexin36 with zonula occludens-1 in HeLa cells, βtC-3 cells, pancreas, and adrenal gland

Xinbo Li, Carl Olson, Shijun Lu, James I. Nagy

Research output: Contribution to journalArticlepeer-review

51 Scopus citations

Abstract

The PDZ domain-containing protein zonula occludens-1 (ZO-1), a well-established component of tight junctions, has recently been shown to interact with various connexin proteins that form gap junctions. We investigated the association of connexin36 (Cx36) with ZO-1 in various cultured cells and tissues. Punctate immunofluorescence labeling for Cx36 was detected in Cx36-transfected HeLa cells, βTC-3 cells, pancreatic islets, and adrenal medulla. Immunofluorescence for ZO-1 was also punctate in cells and tissues, and was colocalized with Cx36 at points of cell-cell contact. Immunoprecipitation of either Cx36 or ZO-1 from cell lysates and tissue homogenates resulted in immunoblot detection of ZO-1 or Cx36, respectively, in immunoprecipitates. A 14-amino acid peptide corresponding to the carboxy-terminus of Cx36 showed binding capacity to the PDZ1 domain of ZO-1, which was eliminated after removal of the last 4 carboxy-terminus amino acids. Low micromolar concentrations of the 14-amino acid peptide produced up to 85% inhibition of Cx36 interaction with the PDZ1 domain of ZO-1. These results provide evidence for molecular interaction between Cx36 and ZO-1 in vitro, and in vivo, and suggest that the interference with Cx36/ZO-1 interaction by short carboxy-terminus peptides of Cx36 may be of value for functional studies of this interaction.

Original languageEnglish (US)
Pages (from-to)485-498
Number of pages14
JournalHistochemistry and Cell Biology
Volume122
Issue number5
DOIs
StatePublished - Nov 2000
Externally publishedYes

Keywords

  • Adrenal medulla
  • Connexin36
  • Gap junctions
  • Pancreas
  • ZO-1

ASJC Scopus subject areas

  • Histology
  • Molecular Biology
  • Medical Laboratory Technology
  • Cell Biology

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