4-Hydroxynonenal induces Cx46 hemichannel inhibition through its carbonylation

Mauricio A. Retamal, Mariana C. Fiori, Ainoa Fernandez-Olivares, Sergio Linsambarth, Francisca Peña, Daisy Quintana, Jimmy Stehberg, Guillermo A. Altenberg

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

Hemichannels formed by connexins mediate the exchange of ions and signaling molecules between the cytoplasm and the extracellular milieu. Under physiological conditions hemichannels have a low open probability, but in certain pathologies their open probability increases, which can result in cell damage. Pathological conditions are characterized by the production of a number of proinflammatory molecules, including 4-hydroxynonenal (4-HNE), one of the most common lipid peroxides produced in response to inflammation and oxidative stress. The aim of this work was to evaluate whether 4-HNE modulates the activity of Cx46 hemichannels. We found that 4-HNE (100 μM) reduced the rate of 4′,6-diamino-2-fenilindol (DAPI) uptake through hemichannels formed by recombinant human Cx46 fused to green fluorescent protein, an inhibition that was reversed partially by 10 mM dithiothreitol. Immunoblot analysis showed that the recombinant Cx46 expressed in HeLa cells becomes carbonylated after exposure to 4-HNE, and that 10 mM dithiothreitol reduced its carbonylation. We also found that Cx46 was carbonylated by 4-HNE in the lens of a selenite-induced cataract animal model. The exposure to 100 μM 4-HNE decreased hemichannel currents formed by recombinant rat Cx46 in Xenopus laevis oocytes. This inhibition also occurred in a mutant expressing only the extracellular loop cysteines, suggesting that other Cys are not responsible for the hemichannel inhibition by carbonylation. This work demonstrates for the first time that Cx46 is post-translationally modified by a lipid peroxide and that this modification reduces Cx46 hemichannel activity.

Original languageEnglish
Article number158705
JournalBiochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
Volume1865
Issue number8
DOIs
Publication statusPublished - Aug 2020

Keywords

  • Carbonylation
  • Cataract
  • Connexin
  • Lipid peroxide
  • Post-translational modification

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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