Ultra-high vacuum surface analysis study of rhodopsin incorporation into supported lipid bilayers

Roger Michel, Varuni Subramaniam, Sally L. McArthur, Bruce Bondurant, Gemma D. D'Ambruoso, Henry K. Hall, Michael F Brown, Eric E. Ross, Steven S Saavedra, David G. Castner

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Planar supported lipid bilayers that are stable under ambient atmospheric and ultra-high-vacuum conditions were prepared by cross-linking polymerization of bis-sorbylphosphatidylcholine (bis-SorbPC). X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) were employed to investigate bilayers that were cross-linked using either redox-initiated radical polymerization or ultraviolet photopolymerization. The redox method yields a more structurally intact bilayer; however, the UV method is more compatible with incorporation of transmembrane proteins. UV polymerization was therefore used to prepare cross-linked bilayers with incorporated bovine rhodopsin, a light-activated, G-protein-coupled receptor (GPCR). A previous study (Subramaniam, V.; Alves, I. D.; Salgado, G. F. J.; Lau, P. W.; Wysocki, R. J.; Salamon, Z.; Tollin, G.; Hruby, V. J.; Brown, M. F.; Saavedra, S. S. J. Am. Chem. Soc. 2005, 127, 5320-5321) showed that rhodopsin retains photoactivity after incorporation into UV-polymerized bis-SorbPC, but did not address how the protein is associated with the bilayer. In this study, we show that rhodopsin is retained in supported bilayers of poly(bis-SorbPC) under ultra-high-vacuum conditions, on the basis of the increase in the XPS nitrogen concentration and the presence of characteristic amino acid peaks in the ToF-SIMS data. Angle-resolved XPS data show that the protein is inserted into the bilayer, rather than adsorbed on the bilayer surface. This is the first study to demonstrate the use of ultra-high-vacuum techniques for structural studies of supported proteolipid bilayers.

Original languageEnglish (US)
Pages (from-to)4901-4906
Number of pages6
JournalLangmuir
Volume24
Issue number9
DOIs
StatePublished - May 6 2008

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Lipid bilayers
Rhodopsin
Surface analysis
Ultrahigh vacuum
ultrahigh vacuum
lipids
X ray photoelectron spectroscopy
Secondary ion mass spectrometry
proteins
Proteins
polymerization
photoelectron spectroscopy
Polymerization
secondary ion mass spectrometry
Proteolipids
Photopolymerization
Free radical polymerization
G-Protein-Coupled Receptors
x rays
Nitrogen

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Colloid and Surface Chemistry

Cite this

Michel, R., Subramaniam, V., McArthur, S. L., Bondurant, B., D'Ambruoso, G. D., Hall, H. K., ... Castner, D. G. (2008). Ultra-high vacuum surface analysis study of rhodopsin incorporation into supported lipid bilayers. Langmuir, 24(9), 4901-4906. https://doi.org/10.1021/la800037r

Ultra-high vacuum surface analysis study of rhodopsin incorporation into supported lipid bilayers. / Michel, Roger; Subramaniam, Varuni; McArthur, Sally L.; Bondurant, Bruce; D'Ambruoso, Gemma D.; Hall, Henry K.; Brown, Michael F; Ross, Eric E.; Saavedra, Steven S; Castner, David G.

In: Langmuir, Vol. 24, No. 9, 06.05.2008, p. 4901-4906.

Research output: Contribution to journalArticle

Michel, R, Subramaniam, V, McArthur, SL, Bondurant, B, D'Ambruoso, GD, Hall, HK, Brown, MF, Ross, EE, Saavedra, SS & Castner, DG 2008, 'Ultra-high vacuum surface analysis study of rhodopsin incorporation into supported lipid bilayers', Langmuir, vol. 24, no. 9, pp. 4901-4906. https://doi.org/10.1021/la800037r
Michel R, Subramaniam V, McArthur SL, Bondurant B, D'Ambruoso GD, Hall HK et al. Ultra-high vacuum surface analysis study of rhodopsin incorporation into supported lipid bilayers. Langmuir. 2008 May 6;24(9):4901-4906. https://doi.org/10.1021/la800037r
Michel, Roger ; Subramaniam, Varuni ; McArthur, Sally L. ; Bondurant, Bruce ; D'Ambruoso, Gemma D. ; Hall, Henry K. ; Brown, Michael F ; Ross, Eric E. ; Saavedra, Steven S ; Castner, David G. / Ultra-high vacuum surface analysis study of rhodopsin incorporation into supported lipid bilayers. In: Langmuir. 2008 ; Vol. 24, No. 9. pp. 4901-4906.
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