Functional Demonstration of Connexin—Protein Binding Using Surface Plasmon Resonance

Heather S. Duffy, Mario Delmar, Wanda Coombs, Steven M. Tafftet, Elliot L. Hertzberg, David C. Spray

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Surface plasmon resonance (SPR) allows examination of protein-protein interactions in real time, from which both binding affinities and kinetics can be directly determined. We have used the SPR technique to search for proteins in heart tissue that would be candidate binding partners for the cardiac gap junction protein, connexin43 (Cx43). Heart lysate showed a strong, pH-dependent binding to the carboxyl terminus (CT) of Cx43 (amino acids 254-382) covalently linked to an SPR cuvette. Binding was inhibited by the presence of v-src transfected 3T3 cell lysate, suggesting that binding partners in these two lysates may compete for overlapping epitopes on Cx43CT. The combined application of proteomic and functional studies is expected to identify which proteins within heart tissue interact with Cx43 and what roles they may play in gap junction function.

Original languageEnglish (US)
Pages (from-to)225-229
Number of pages5
JournalCell Communication and Adhesion
Volume8
Issue number42100
DOIs
Publication statusPublished - Jan 1 2001

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Keywords

  • Connexin43
  • ZO-1
  • connexin-protein
  • surface plasmon resonance

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology

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