title: G-protein-coupled receptors in drug discovery: nanosizing using cell-free technologies and molecular biology approaches.
abstract: Signal transduction by G-protein-coupled receptors (GPCRs) underpins a multitude of physiological processes. Ligand recognition by the receptor leads to activation of a generic molecular switch involving heterotrimeric G-proteins and guanine nucleotides. Signal transduction has been studied extensively with both cell-based systems and assays comprising isolated signaling components. Interest and commercial investment in GPCRs in areas such as drug targets, orphan receptors, high throughput screening, biosensors, and so on will focus greater attention on assay development to allow for miniaturization, ultra-high throughput and, eventually, microarray/biochip assay formats. Although cell-based assays are adequate for many GPCRs, it is likely that these formats will limit the development of higher density GPCR assay platforms mandatory for other applications. Stable, robust, cell-free signaling assemblies comprising receptor and appropriate molecular switching components will form the basis of future GPCR assay platforms adaptable for such applications as microarrays. The authors review current cell-free GPCR assay technologies and molecular biological approaches for construction of novel, functional GPCR assays.
authors.affiliation: CSIRO Molecular and Health Technologies, Adelaide, SA, Australia. wayne.leifert@csiro.au
authors.forename: Wayne R
authors.initials: WR
authors.lastname: Leifert
authors.forename: Amanda L
authors.initials: AL
authors.lastname: Aloia
authors.forename: Olgatina
authors.initials: O
authors.lastname: Bucco
authors.forename: Richard V
authors.initials: RV
authors.lastname: Glatz
authors.forename: Edward J
authors.initials: EJ
authors.lastname: McMurchie
mesh_terms: D001681:Biological Assay; D015374:Biosensing Techniques; D002474:Cell-Free System; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006160:Guanosine Triphosphate / Q000378:metabolism*; D008967:Molecular Biology; D036103:Nanotechnology; D040081:Protein Array Analysis; D015202:Protein Engineering; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D043562:Receptors, G-Protein-Coupled; D006160:Guanosine Triphosphate