Kinetics of protein-protein interactions of connexins: Use of enzyme linked sorbent assays

Heather S. Duffy, Phyllis O'Donnell, Wanda Coombs, Steven M. Taffet, Mario Delmar, David C. Spray

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Determination of the protein-protein interactions of connexins has become a rapidly expanding field of research. While there are multiple methods of determining the identity of binding partners, determination of the strengths of interactions is not as simple. Here we describe the use of the in vitro method Enzyme Linked Sorbent Assay (ELSA) to compare binding affinities of known protein partners for Connexin43. We used the binding of Cx43 Carboxyl Terminal domain to the PDZ-2 domain of Zonula Occludens-1 and to the SH3 domain of c-Src. In the ELSA assay we found that while the binding of the SH3 domain of c-Src is pH-dependent, the interaction of the PDZ domain of ZO-1 is not. These data confirm findings using Surface Plasmon Resonance (1) and indicate that ELSA can be a useful tool in determining the kinetics of protein-protein interactions.

Original languageEnglish (US)
Pages (from-to)207-210
Number of pages4
JournalCell Communication and Adhesion
Volume10
Issue number4-6
StatePublished - Jul 2003

Fingerprint

Connexins
Sorbents
Assays
Kinetics
PDZ Domains
Enzymes
Connexin 43
src Homology Domains
Proteins
Surface Plasmon Resonance
Tight Junctions
Surface plasmon resonance
Research

Keywords

  • Connexin-protein interactions
  • Connexins
  • Cx43
  • Gap junctions
  • Nexus

ASJC Scopus subject areas

  • Cell Biology
  • Clinical Biochemistry

Cite this

Duffy, H. S., O'Donnell, P., Coombs, W., Taffet, S. M., Delmar, M., & Spray, D. C. (2003). Kinetics of protein-protein interactions of connexins: Use of enzyme linked sorbent assays. Cell Communication and Adhesion, 10(4-6), 207-210.

Kinetics of protein-protein interactions of connexins : Use of enzyme linked sorbent assays. / Duffy, Heather S.; O'Donnell, Phyllis; Coombs, Wanda; Taffet, Steven M.; Delmar, Mario; Spray, David C.

In: Cell Communication and Adhesion, Vol. 10, No. 4-6, 07.2003, p. 207-210.

Research output: Contribution to journalArticle

Duffy, HS, O'Donnell, P, Coombs, W, Taffet, SM, Delmar, M & Spray, DC 2003, 'Kinetics of protein-protein interactions of connexins: Use of enzyme linked sorbent assays', Cell Communication and Adhesion, vol. 10, no. 4-6, pp. 207-210.
Duffy, Heather S. ; O'Donnell, Phyllis ; Coombs, Wanda ; Taffet, Steven M. ; Delmar, Mario ; Spray, David C. / Kinetics of protein-protein interactions of connexins : Use of enzyme linked sorbent assays. In: Cell Communication and Adhesion. 2003 ; Vol. 10, No. 4-6. pp. 207-210.
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