Rhodopsin reconstituted into a planar-supported lipid bilayer retains photoactivity after cross-linking polymerization of lipid monomers.
Transmembrane proteins (TMPs), particularly ion channels and receptors, play key roles in transport and signal transduction. Many of these proteins are pharmacologically important and therefore targets for drug discovery. TMPs can be reconstituted in planar-supported lipid bilayers (PSLBs), which has led to development of TMP-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 TMP structure and activity. In this study, we have examined the effects of UV polymerization of bis-Sorbylphosphatidylcholine (bis-SorbPC) on the photoactivation of reconstituted bovine rhodopsin (Rho), a model G-protein-coupled receptor. Plasmon-waveguide resonance spectroscopy (PWR) was used to compare the degree of Rho incorporation and activation in fluid and poly(lipid) PSLBs. The results show that reconstitution of Rho into a supported lipid bilayer composed only of bis-SorbPC, followed by photoinduced lipid cross-linking, does not measurably diminish protein function.
Subramaniam
Varuni
V
Department of Chemistry, University of Arizona, Tucson, Arizona, 85721-0041, USA.
Alves
Isabel D
ID
Salgado
Gilmar F J
GF
Lau
Pick-Wei
PW
Wysocki
Ronald J
RJ
Salamon
Zdzislaw
Z
Tollin
Gordon
G
Hruby
Victor J
VJ
Brown
Michael F
MF
Saavedra
S Scott
SS
D015374:Biosensing Techniques / Q000379:methods; D003432:Cross-Linking Reagents / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry; D010777:Photochemistry; D012243:Rhodopsin / Q000737:chemistry*; D020349:Surface Plasmon Resonance; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; C077390:1,2-bis(10(2',4'-hexadienoyloxy)decanoyl)-sn-glycero-3-phosphocholine; D012243:Rhodopsin