Assembly of purple membranes on polyelectrolyte films

Marie Belle Saab, Elias Estephan, Thierry Cloitre, René Legros, FrédéricJGCuisinier, László Zimányi, Csilla Gergely

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The membrane protein bacteriorhodopsin in its native membrane bound form (purple membrane) was adsorbed and incorporated into polyelectrolyte multilayered films, and adsorption was in situ monitored by optical waveguide light- mode spectroscopy. The formation of a single layer or a double layer of purple membranes was observed when adsorbed on negatively or positively charged surfaces, respectively. The purple membrane patches adsorbed on the polyelectrolyte multilayers were also evidenced by atomic force microscopy images. The driving forces of the adsorption process were evaluated by varying the ionic strength of the solution as well as the purple membrane concentration. At high purple membrane concentration, interpenetrating polyelectrolyte loops might provide new binding sites for the adsorption of a second layer of purple membranes, whereas at lower concentrations only a single layer is formed. Negative surfaces do not promote a second protein layer adsorption. Driving forces other than just electrostatic ones, such as hydrophobic forces, should play a role in the polyelectrolyte/purple membrane layering. The subtle interplay of all these factors determines the formation of the polyelectrolyte/purple membrane matrix with a presumably high degree of orientation for the incorporated purple membranes, with their cytoplasmic, or extracellular side toward the bulk on negatively or positively charged polyelectrolyte, respectively. The structural stability of bacteriorhodopsin during adsorption onto the surface and incorporation into the polyelectrolyte multilayers was investigated by Fourier transform infrared spectro- scopy in attenuated total reflection mode. Adsorption and incorporation of purple membranes within polyelectrolyte multilayers does not disturb the conformational majority of membrane-embedded α-helix structures of the protein, but may slightly alter the structure of the extramembraneous segments or their interaction with the environment. This high stability is different from the lower stability of the predominantly β-sheet structures of numerous globular proteins when adsorbed onto surfaces.

Original languageEnglish
Pages (from-to)5159-5167
Number of pages9
JournalLangmuir
Volume25
Issue number9
DOIs
Publication statusPublished - May 5 2009

ASJC Scopus subject areas

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

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    Saab, M. B., Estephan, E., Cloitre, T., Legros, R., FrédéricJGCuisinier, Zimányi, L., & Gergely, C. (2009). Assembly of purple membranes on polyelectrolyte films. Langmuir, 25(9), 5159-5167. https://doi.org/10.1021/la9002274