| Ultra-high vacuum surface analysis study of rhodopsin incorporation into supported lipid bilayers. | |
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MedLine Citation:
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PMID: 18393486 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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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:
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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:
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Title: Langmuir : the ACS journal of surfaces and colloids Volume: 24 ISSN: 0743-7463 ISO Abbreviation: Langmuir Publication Date: 2008 May |
Date Detail:
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Created Date: 2008-04-30 Completed Date: 2008-06-11 Revised Date: 2013-04-01 |
Medline Journal Info:
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Nlm Unique ID: 9882736 Medline TA: Langmuir Country: United States |
Other Details:
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Languages: eng Pagination: 4901-6 Citation Subset: IM |
Affiliation:
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National ESCA and Surface Analysis Center for Biomedical Problems, Department of Bioengineering, Box 351750, University of Washington, Seattle, Washington 98195, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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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:
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EB-002027/EB/NIBIB NIH HHS; EB007047/EB/NIBIB NIH HHS; EY 12049/EY/NEI 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:
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0/Lipid Bilayers; 0/Phosphatidylcholines; 0/Polymers; 7440-44-0/Carbon; 9009-81-8/Rhodopsin |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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