Effects of phosphorylation on the structure and backbone dynamics of the intrinsically disordered connexin43 C-terminal domain

Rosslyn Grosely, Jennifer L. Kopanic, Sarah Nabors, Fabien Kieken, Gaëlle Spagnol, Mona Al-Mugotir, Sydney Zach, Paul L Sorgen

Research output: Contribution to journalArticle

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Abstract

Background: Phosphorylation of the connexin43 C-terminal (Cx43CT) domain regulates gap junction intercellular communication (GJIC). Results: Phosphorylation alters the α-helical propensity of the Cx43CT. Conclusion: Altering the conformational preference of the Cx43CT presents a novel mechanism for regulation of GJIC. Significance: Cx43CT residues susceptible to structural alterations are prime targets for chemical modulators of GJIC. Phosphorylation of the connexin43 C-terminal (Cx43CT) domain regulates gap junction intercellular communication. However, an understanding of the mechanisms by which phosphorylation exerts its effects is lacking. Here, we test the hypothesis that phosphorylation regulates Cx43 gap junction intercellular communication by mediating structural changes in the C-terminal domain. Circular dichroism and nuclear magnetic resonance were used to characterize the effects of phosphorylation on the secondary structure and backbone dynamics of soluble and membrane-tethered Cx43CT domains. Cx43CT phospho-mimetic isoforms, which have Asp substitutions at specific Ser/Tyr sites, revealed phosphorylation alters the α-helical content of the Cx43CT domain only when attached to the membrane. The changes in secondary structure are due to variations in the conformational preference and backbone flexibility of residues adjacent and distal to the site(s) of modification. In addition to the known direct effects of phosphorylation on molecular partner interactions, the data presented here suggest phosphorylation may also indirectly regulate binding affinity by altering the conformational preference of the Cx43CT domain.

Original languageEnglish (US)
Pages (from-to)24857-24870
Number of pages14
JournalJournal of Biological Chemistry
Volume288
Issue number34
DOIs
StatePublished - Aug 23 2013

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Connexin 43
Phosphorylation
Gap Junctions
Communication
Membranes
Molecular interactions
Viperidae
Circular Dichroism
Modulators
Protein Isoforms
Substitution reactions
Magnetic Resonance Spectroscopy
Nuclear magnetic resonance

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Effects of phosphorylation on the structure and backbone dynamics of the intrinsically disordered connexin43 C-terminal domain. / Grosely, Rosslyn; Kopanic, Jennifer L.; Nabors, Sarah; Kieken, Fabien; Spagnol, Gaëlle; Al-Mugotir, Mona; Zach, Sydney; Sorgen, Paul L.

In: Journal of Biological Chemistry, Vol. 288, No. 34, 23.08.2013, p. 24857-24870.

Research output: Contribution to journalArticle

Grosely, Rosslyn ; Kopanic, Jennifer L. ; Nabors, Sarah ; Kieken, Fabien ; Spagnol, Gaëlle ; Al-Mugotir, Mona ; Zach, Sydney ; Sorgen, Paul L. / Effects of phosphorylation on the structure and backbone dynamics of the intrinsically disordered connexin43 C-terminal domain. In: Journal of Biological Chemistry. 2013 ; Vol. 288, No. 34. pp. 24857-24870.
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