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G‐protein coupled receptor array technologies: Site directed immobilisation of liposomes containing the H 1 ‐histamine or M 2 ‐muscarinic receptors
Author(s) -
Bailey Kelly,
Bally Marta,
Leifert Wayne,
Vörös Janos,
McMurchie Ted
Publication year - 2009
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200800539
Subject(s) - muscarinic acetylcholine receptor , histamine receptor , histamine , receptor , histamine h4 receptor , liposome , chemistry , g protein coupled receptor , muscarinic acetylcholine receptor m5 , histamine h1 receptor , pharmacology , biochemistry , biology , muscarinic acetylcholine receptor m3 , histamine h2 receptor , antagonist
This paper describes a novel strategy to create a microarray of G‐protein coupled receptors (GPCRs), an important group of membrane proteins both physiologically and pharmacologically. The H 1 ‐histamine receptor and the M 2 ‐muscarinic receptor were both used as model GPCRs in this study. The receptor proteins were embedded in liposomes created from the cellular membrane extracts of Spodoptera frugiperda ( Sf9 ) insect cell culture line with its accompanying baculovirus protein insert used for overexpression of the receptors. Once captured onto a surface these liposomes provide a favourable lipidic environment for the integral membrane proteins. Site directed immobilisation of these liposomes was achieved by introduction of cholesterol‐modified oligonucleotides (oligos). These oligo/cholesterol conjugates incorporate within the lipid bilayer and were captured by the complementary oligo strand exposed on the surface. Sequence specific immobilisation was demonstrated using a quartz crystal microbalance with dissipation (QCM‐D). Confirmatory results were also obtained by monitoring fluorescent ligand binding to GPCRs captured on a spotted oligo microarray using Confocal Laser Scanning Microscopy and the ZeptoREADER microarray imaging system. Sequence specific immobilisation of such biologically important membrane proteins could lead to the development of a heterogeneous self‐sorting liposome array of GPCRs which would underpin a variety of future novel applications.