Reconstitution of rhodopsin into polymerizable planar supported lipid bilayers : influence of dienoyl monomer structure on photoactivation

G-protein-coupled receptors (GPCRs) play key roles in cellular signal transduction and many are pharmacologically important targets for drug discovery. GPCRs can be reconstituted in planar supported lipid bilayers (PSLBs) with retention of activity, which has led to development of GPCR-based biosens...

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Veröffentlicht in:Langmuir : the ACS journal of surfaces and colloids. - 1999. - 24(2008), 19 vom: 07. Okt., Seite 11067-75
1. Verfasser: Subramaniam, Varuni (VerfasserIn)
Weitere Verfasser: D'Ambruoso, Gemma D, Hall, H K Jr, Wysocki, Ronald J Jr, Brown, Michael F, Saavedra, S Scott
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2008
Zugriff auf das übergeordnete Werk:Langmuir : the ACS journal of surfaces and colloids
Schlagworte:Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S. Lipid Bilayers Polymers Octoxynol 9002-93-1 Rhodopsin 9009-81-8
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100 1 |a Subramaniam, Varuni  |e verfasserin  |4 aut 
245 1 0 |a Reconstitution of rhodopsin into polymerizable planar supported lipid bilayers  |b influence of dienoyl monomer structure on photoactivation 
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520 |a G-protein-coupled receptors (GPCRs) play key roles in cellular signal transduction and many are pharmacologically important targets for drug discovery. GPCRs can be reconstituted in planar supported lipid bilayers (PSLBs) with retention of activity, which has led to development of GPCR-based biosensors and biochips. However, PSLBs composed of natural lipids lack the high stability desired for many technological applications. One strategy is to use synthetic lipid monomers that can be polymerized to form robust bilayers. A key question is how lipid polymerization affects GPCR structure and activity. Here we have investigated the photochemical activity of bovine rhodopsin (Rho), a model GPCR, reconstituted into PSLBs composed of lipids having one or two polymerizable dienoyl moieties located in different regions of the acyl chains. Plasmon waveguide resonance spectroscopy was used to compare the degree of Rho photoactivation in fluid and poly(lipid) PSLBs. The position of the dienoyl moiety was found to have a significant effect: polymerization near the glycerol backbone significantly attenuates Rho activity whereas polymerization near the acyl chain termini does not. Differences in cross-link density near the acyl chain termini also do not affect Rho activity. In unpolymerized PSLBs, an equimolar mixture of phosphatidylethanolamine and phosphatidylcholine (PC) lipids enhances activity relative to pure PC; however after polymerization, the enhancement is eliminated which is attributed to stabilization of the membrane lamellar phase. These results should provide guidance for the design of robust lipid bilayers functionalized with transmembrane proteins for use in membrane-based biochips and biosensors 
650 4 |a Journal Article 
650 4 |a Research Support, N.I.H., Extramural 
650 4 |a Research Support, U.S. Gov't, Non-P.H.S. 
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650 7 |a Polymers  |2 NLM 
650 7 |a Octoxynol  |2 NLM 
650 7 |a 9002-93-1  |2 NLM 
650 7 |a Rhodopsin  |2 NLM 
650 7 |a 9009-81-8  |2 NLM 
700 1 |a D'Ambruoso, Gemma D  |e verfasserin  |4 aut 
700 1 |a Hall, H K  |c Jr  |e verfasserin  |4 aut 
700 1 |a Wysocki, Ronald J  |c Jr  |e verfasserin  |4 aut 
700 1 |a Brown, Michael F  |e verfasserin  |4 aut 
700 1 |a Saavedra, S Scott  |e verfasserin  |4 aut 
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773 1 8 |g volume:24  |g year:2008  |g number:19  |g day:07  |g month:10  |g pages:11067-75 
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