title: Fragment screening by SPR and advanced application to GPCRs.
abstract: Surface plasmon resonance (SPR) is one of the primary biophysical methods for the screening of low molecular weight 'fragment' libraries, due to its low protein consumption and 'label-free' methodology. SPR biosensor interaction analysis is employed to both screen and confirm the binding of compounds in fragment screening experiments, as it provides accurate information on the affinity and kinetics of molecular interactions. The most advanced application of the use of SPR for fragment screening is against membrane protein drug targets, such G-protein coupled receptors (GPCRs). Biophysical GPCR assays using SPR have been validated with pharmacological measurements approximate to cell-based methods, yet provide the advantage of biophysical methods in their ability to measure the weak affinities of low molecular weight fragments. A number of SPR fragment screens against GPCRs have now been disclosed in the literature. SPR fragment screening is proving versatile to screen both thermostabilised GPCRs and solubilised wild type receptors. In this chapter, we discuss the state-of-the-art in GPCR fragment screening by SPR and the technical considerations in performing such experiments.
authors.affiliation: Division of Biological Chemistry and Drug Discovery, College of Life Science, University of Dundee, Dow Street, Dundee DD1 5EH, United Kingdom.
authors.forename: Claire A
authors.initials: CA
authors.lastname: Shepherd
authors.affiliation: Division of Biological Chemistry and Drug Discovery, College of Life Science, University of Dundee, Dow Street, Dundee DD1 5EH, United Kingdom. Electronic address: a.hopkins@dundee.ac.uk.
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Hopkins
authors.affiliation: Division of Biological Chemistry and Drug Discovery, College of Life Science, University of Dundee, Dow Street, Dundee DD1 5EH, United Kingdom.
authors.forename: Iva
authors.initials: I
authors.lastname: Navratilova
mesh_terms: D000818:Animals; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D055550:Protein Stability; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D043562:Receptors, G-Protein-Coupled
keywords: Biophysical screening; Fragment screening; G-protein coupled receptors; GPCRs; Membrane proteins; SPR; Surface plasmon resonance