title: Frontal affinity chromatography-mass spectrometry useful for characterization of new ligands for GPR17 receptor.
abstract: The application of frontal affinity chromatography-mass spectrometry (FAC-MS), along with molecular modeling studies, to the screening of potential drug candidates toward the recently deorphanized G-protein-coupled receptor (GPCR) GPR17 is shown. GPR17 is dually activated by uracil nucleotides and cysteinyl-leukotrienes, and is expressed in organs typically undergoing ischemic damage (i.e., brain, heart and kidney), thus representing a new pharmacological target for acute and chronic neurodegeneration. GPR17 was entrapped on an immobilized artificial membrane (IAM), and this stationary phase was used to screen a library of nucleotide derivatives by FAC-MS to select high affinity ligands. The chromatographic results have been validated with a reference functional assay ([(35)S]GTPgammaS binding assay). The receptor nucleotide-binding site was studied by setting up a column where a mutated GPR17 receptor (Arg255Ile) has been immobilized. The chromatographic behavior of the tested nucleotide derivatives together with in silico studies have been used to gain insights into the structure requirement of GPR17 ligands.
authors.affiliation: Department of Pharmaceutical Chemistry, University of Pavia, Via Taramelli 12, 27100 Pavia, Italy.
authors.forename: Enrica
authors.initials: E
authors.lastname: Calleri
authors.forename: Stefania
authors.initials: S
authors.lastname: Ceruti
authors.forename: Gloria
authors.initials: G
authors.lastname: Cristalli
authors.forename: Claudia
authors.initials: C
authors.lastname: Martini
authors.forename: Caterina
authors.initials: C
authors.lastname: Temporini
authors.forename: Chiara
authors.initials: C
authors.lastname: Parravicini
authors.forename: Rosaria
authors.initials: R
authors.lastname: Volpini
authors.forename: Simona
authors.initials: S
authors.lastname: Daniele
authors.forename: Gabriele
authors.initials: G
authors.lastname: Caccialanza
authors.forename: Davide
authors.initials: D
authors.lastname: Lecca
authors.forename: Catia
authors.initials: C
authors.lastname: Lambertucci
authors.forename: Maria Letizia
authors.initials: ML
authors.lastname: Trincavelli
authors.forename: Gabriella
authors.initials: G
authors.lastname: Marucci
authors.forename: Irving W
authors.initials: IW
authors.lastname: Wainer
authors.forename: Graziella
authors.initials: G
authors.lastname: Ranghino
authors.forename: Piercarlo
authors.initials: P
authors.lastname: Fantucci
authors.forename: Maria P
authors.initials: MP
authors.lastname: Abbracchio
authors.forename: Gabriella
authors.initials: G
authors.lastname: Massolini
mesh_terms: D001665:Binding Sites; D002846:Chromatography, Affinity; D006801:Humans; D008024:Ligands; D013058:Mass Spectrometry; D008567:Membranes, Artificial; D008958:Models, Molecular; D009711:Nucleotides; D011485:Protein Binding; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: C121670:GPR17 protein, human; D008024:Ligands; D008567:Membranes, Artificial; D009711:Nucleotides; D043562:Receptors, G-Protein-Coupled
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