title: Techniques: GPCR assembly, pharmacology and screening by flow cytometry.
abstract: Flow cytometers are well known for their ability to analyze and sort cells at high rates based on physiological responses and expression of protein markers. The potential for flow cytometry in G-protein-coupled receptor (GPCR) research, however, is less well appreciated. Potential applications include: (i) the homogenous discrimination of free and bound ligands or proteins in both cellular and microsphere-based assays; and (ii) multiplexed ('suspension array') analysis of cell responses and protein-protein interactions. Innovative sample-handling systems also provide sub-second resolution of interaction kinetics and 1 second per well throughput of microliter-sized samples from multiwell plates. Flow cytometric methods using microspheres for analysis of GPCRs that interact with intracellular and extracellular binding partners such as ligands, G proteins and kinases have been established. These analyses can produce quantitative pharmacological data analogous to radioligand assays, and, in some cases, the probes can be integrated into the assembly as fluorescent fusion proteins.
authors.affiliation: Department of Pathology and Cancer Center, University of New Mexico Health Science Center, Albuquerque, NM 87131, USA.
authors.forename: Anna
authors.initials: A
authors.lastname: Waller
authors.forename: Peter C
authors.initials: PC
authors.lastname: Simons
authors.forename: Sean M
authors.initials: SM
authors.lastname: Biggs
authors.forename: Bruce S
authors.initials: BS
authors.lastname: Edwards
authors.forename: Eric R
authors.initials: ER
authors.lastname: Prossnitz
authors.forename: Larry A
authors.initials: LA
authors.lastname: Sklar
mesh_terms: D000818:Animals; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008024:Ligands; D043562:Receptors, G-Protein-Coupled / Q000032:analysis* / Q000187:drug effects* / Q000502:physiology; D015398:Signal Transduction
chemical_list: D008024:Ligands; D043562:Receptors, G-Protein-Coupled