Effect of the microenvironment on the recognition of immobilized cytochromes by soluble redox proteins

A. A. Kloss, N. Lavrik, C. Yeung, Deborah E Leckband

Research output: Contribution to journalArticle

Abstract

This work examines the effect of the electrostatic properties of an immobilization substrate on the apparent affinity of immobilized receptors for their soluble ligands. In particular, we report measurements of the interaction between cytochrome c or cytochrome b5 bound to an oriented streptavidin monolayer and its soluble reaction partner. Surface plasmon resonance measurements demonstrate that the charge on the underlying streptavidin monolayer has a pronounced impact on the measured equilibrium constants for the heterologous interaction between cyt c and cyt b5. The pH dependence of the equilibrium binding constant is similar qualitatively to that reported for the protein interaction in solution. However, the pH optima of the affinities measured with the immobilized proteins are shifted by up to 1.0 pH units. Whether the shift is to acidic or basic pH depends on which of the two proteins is immobilized. The observed changes are consistent with the long-range repulsion or attraction between the soluble ligand and the supporting streptavidin monolayer.

Original languageEnglish (US)
Pages (from-to)3414-3421
Number of pages8
JournalLangmuir
Volume16
Issue number7
DOIs
StatePublished - Apr 4 2000

Fingerprint

Streptavidin
cytochromes
Cytochromes
Monolayers
proteins
Proteins
Immobilized Proteins
Ligands
affinity
Cytochromes b5
Equilibrium constants
Surface plasmon resonance
Cytochromes c
ligands
Electrostatics
interactions
immobilization
surface plasmon resonance
attraction
Substrates

ASJC Scopus subject areas

  • Materials Science(all)
  • Condensed Matter Physics
  • Surfaces and Interfaces
  • Spectroscopy
  • Electrochemistry

Cite this

Effect of the microenvironment on the recognition of immobilized cytochromes by soluble redox proteins. / Kloss, A. A.; Lavrik, N.; Yeung, C.; Leckband, Deborah E.

In: Langmuir, Vol. 16, No. 7, 04.04.2000, p. 3414-3421.

Research output: Contribution to journalArticle

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