Document Detail


Ultra-high vacuum surface analysis study of rhodopsin incorporation into supported lipid bilayers.
MedLine Citation:
PMID:  18393486     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Roger Michel; Varuni Subramaniam; Sally L McArthur; Bruce Bondurant; Gemma D D'Ambruoso; Henry K Hall; Michael F Brown; Eric E Ross; S Scott Saavedra; David G Castner
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2008-04-05
Journal Detail:
Title:  Langmuir : the ACS journal of surfaces and colloids     Volume:  24     ISSN:  0743-7463     ISO Abbreviation:  Langmuir     Publication Date:  2008 May 
Date Detail:
Created Date:  2008-04-30     Completed Date:  2008-06-11     Revised Date:  2013-11-12    
Medline Journal Info:
Nlm Unique ID:  9882736     Medline TA:  Langmuir     Country:  United States    
Other Details:
Languages:  eng     Pagination:  4901-6     Citation Subset:  IM    
Affiliation:
National ESCA and Surface Analysis Center for Biomedical Problems, Department of Bioengineering, Box 351750, University of Washington, Seattle, Washington 98195, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Carbon / chemistry
Cattle
Lipid Bilayers / chemistry*
Mass Spectrometry
Oxidation-Reduction
Phosphatidylcholines / chemistry*
Polymers / chemistry
Rhodopsin / chemistry*
Spectrophotometry
Surface Properties
Vacuum
Grant Support
ID/Acronym/Agency:
EB-002027/EB/NIBIB NIH HHS; EB007047/EB/NIBIB NIH HHS; EY 12049/EY/NEI NIH HHS; P41 EB002027/EB/NIBIB NIH HHS; R01 EB007047/EB/NIBIB NIH HHS; R01 EB007047-01/EB/NIBIB NIH HHS; R01 EB007047-02/EB/NIBIB NIH HHS
Chemical
Reg. No./Substance:
0/Lipid Bilayers; 0/Phosphatidylcholines; 0/Polymers; 7440-44-0/Carbon; 9009-81-8/Rhodopsin
Comments/Corrections

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