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

Varuni Subramaniam, Gemma D. D'Ambruoso, H. K. Hall, Ronald J. Wysocki, Michael F Brown, Steven S Saavedra

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

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 (Rhô), 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.

Original languageEnglish (US)
Pages (from-to)11067-11075
Number of pages9
JournalLangmuir
Volume24
Issue number19
DOIs
StatePublished - Oct 7 2008

Fingerprint

Lipid bilayers
Rhodopsin
Lipids
lipids
G-Protein-Coupled Receptors
monomers
Monomers
Proteins
Polymerization
Biochips
proteins
Phosphatidylcholines
Biosensors
polymerization
Membranes
Signal transduction
Glycerol
bioinstrumentation
Waveguides
Thermodynamic properties

ASJC Scopus subject areas

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

Cite this

Reconstitution of rhodopsin into polymerizable planar supported lipid bilayers : Influence of dienoyl monomer structure on photoactivation. / Subramaniam, Varuni; D'Ambruoso, Gemma D.; Hall, H. K.; Wysocki, Ronald J.; Brown, Michael F; Saavedra, Steven S.

In: Langmuir, Vol. 24, No. 19, 07.10.2008, p. 11067-11075.

Research output: Contribution to journalArticle

Subramaniam, Varuni ; D'Ambruoso, Gemma D. ; Hall, H. K. ; Wysocki, Ronald J. ; Brown, Michael F ; Saavedra, Steven S. / Reconstitution of rhodopsin into polymerizable planar supported lipid bilayers : Influence of dienoyl monomer structure on photoactivation. In: Langmuir. 2008 ; Vol. 24, No. 19. pp. 11067-11075.
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