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

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 Adhesion and Communication
Volume8
Issue number4-6
StatePublished - 2000

Fingerprint

Connexins
Surface Plasmon Resonance
Surface plasmon resonance
Protein Binding
Connexin 43
Demonstrations
Proteins
Tissue
3T3 Cells
Gap Junctions
Proteomics
Epitopes
Amino Acids
Kinetics

Keywords

  • Connexin-protein
  • Connexin43
  • Surface plasmon resonance
  • ZO-1

ASJC Scopus subject areas

  • Cell Biology

Cite this

Duffy, H. S., Delmar, M., Coombs, W., Tafftet, S. M., Hertzberg, E. L., & Spray, D. C. (2000). Functional demonstration of connexin-protein binding using surface plasmon resonance. Cell Adhesion and Communication, 8(4-6), 225-229.

Functional demonstration of connexin-protein binding using surface plasmon resonance. / Duffy, Heather S.; Delmar, Mario; Coombs, Wanda; Tafftet, Steven M.; Hertzberg, Elliot L.; Spray, David C.

In: Cell Adhesion and Communication, Vol. 8, No. 4-6, 2000, p. 225-229.

Research output: Contribution to journalArticle

Duffy, HS, Delmar, M, Coombs, W, Tafftet, SM, Hertzberg, EL & Spray, DC 2000, 'Functional demonstration of connexin-protein binding using surface plasmon resonance', Cell Adhesion and Communication, vol. 8, no. 4-6, pp. 225-229.
Duffy, Heather S. ; Delmar, Mario ; Coombs, Wanda ; Tafftet, Steven M. ; Hertzberg, Elliot L. ; Spray, David C. / Functional demonstration of connexin-protein binding using surface plasmon resonance. In: Cell Adhesion and Communication. 2000 ; Vol. 8, No. 4-6. pp. 225-229.
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