Optical anisotropy of flagellin layers: In situ and label-free measurement of adsorbed protein orientation using OWLS

Noemi Kovacs, Daniel Patko, Norbert Orgovan, Sandor Kurunczi, Jeremy J. Ramsden, F. Vonderviszt, R. Horváth

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

The surface adsorption of the protein flagellin was followed in situ using optical waveguide lightmode spectroscopy (OWLS). Flagellin did not show significant adsorption on a hydrophilic waveguide, but very rapidly formed a dense monolayer on a hydrophobic (silanized) surface. The homogeneous and isotropic optical layer model, which has hitherto been generally applied in OWLS data interpretation for adsorbed protein films, failed to characterize the flagellin layer, but it could be successfully modeled as an uniaxial thin film. This anisotropic modeling revealed a significant positive birefringence in the layer, suggesting oriented protein adsorption. The adsorbed flagellin orientation was further evidenced by monitoring the surface adsorption of truncated flagellin variants, in which the terminal protein regions or the central (D3) domain were removed. Without the terminal regions the protein adsorption was much slower and the resulting films were significantly less birefringent, implying that intact flagellin adsorbs on the hydrophobic surface via its terminal regions.

Original languageEnglish
Pages (from-to)5382-5389
Number of pages8
JournalAnalytical Chemistry
Volume85
Issue number11
DOIs
Publication statusPublished - Jun 4 2013

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Flagellin
Optical anisotropy
Optical waveguides
Labels
Spectroscopy
Adsorption
Proteins
Birefringence
Monolayers
Waveguides
Thin films
Monitoring

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Optical anisotropy of flagellin layers : In situ and label-free measurement of adsorbed protein orientation using OWLS. / Kovacs, Noemi; Patko, Daniel; Orgovan, Norbert; Kurunczi, Sandor; Ramsden, Jeremy J.; Vonderviszt, F.; Horváth, R.

In: Analytical Chemistry, Vol. 85, No. 11, 04.06.2013, p. 5382-5389.

Research output: Contribution to journalArticle

Kovacs, Noemi ; Patko, Daniel ; Orgovan, Norbert ; Kurunczi, Sandor ; Ramsden, Jeremy J. ; Vonderviszt, F. ; Horváth, R. / Optical anisotropy of flagellin layers : In situ and label-free measurement of adsorbed protein orientation using OWLS. In: Analytical Chemistry. 2013 ; Vol. 85, No. 11. pp. 5382-5389.
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