Use of surface plasmon resonance for real-time analysis of the interaction of ZO-1 and occludin

Anke Schmidt, Darkhan I. Utepbergenov, Gerd Krause, Ingolf E. Blasig

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Surface plasmon resonance (SPR) spectroscopy was applied to study in real time, the interaction between the tight junction proteins ZO-1 and occludin. To imitate the morphology of tight junctions, a cytosolic tail of mouse occludin was immobilised at the sensor and guanylate kinase-like domain (Guk) was allowed to pass over the modified chip surface. The Guk domain of ZO-1 (residues 644-812) was found to bind to the cytoplasmic, carboxy-terminal region of occludin (residues 378-521). This interaction was systematically characterised with respect to the concentrations of both proteins and the binding conditions. Under the given experimental conditions, association and dissociation showed saturation kinetics, with affinity in micromolar range: ka = 4.14 ± 0.52 × 103 M-1 s-1, kd = 3.04 ± 0.38 × 10-3 s-1, KD = 639 ± 51 nM. The results support the hypothesis that the Guk domain of ZO-1 is involved in the recruitment of the transmembrane protein occludin at tight junctions by interacting with the cytosolic carboxy-terminal sequence of occludin, located far from the cell membrane. We demonstrate the use of SPR spectroscopy as an effective approach for characterisation of the interactions of junction proteins.

Original languageEnglish
Pages (from-to)1194-1199
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume288
Issue number5
DOIs
Publication statusPublished - Nov 16 2001
Externally publishedYes

Fingerprint

Occludin
Surface Plasmon Resonance
Surface plasmon resonance
Guanylate Kinases
Tight Junctions
Spectrum Analysis
Zonula Occludens-1 Protein
Spectroscopy
Proteins
Cell membranes
Protein Binding
Tail
Cell Membrane
Association reactions
Kinetics
Sensors

Keywords

  • Guk domain
  • Occludin
  • Protein interaction
  • Surface plasmon resonance
  • Tight junctions
  • ZO-1

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Use of surface plasmon resonance for real-time analysis of the interaction of ZO-1 and occludin. / Schmidt, Anke; Utepbergenov, Darkhan I.; Krause, Gerd; Blasig, Ingolf E.

In: Biochemical and Biophysical Research Communications, Vol. 288, No. 5, 16.11.2001, p. 1194-1199.

Research output: Contribution to journalArticle

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