title: Development of a CCK1R-membrane nanoparticle as a fish-out tool for bioactive peptides.
abstract: The cholecystokinin receptor type 1 (CCK1R) is a G protein-coupled receptor (GPCR) that is involved in several biological processes including the regulation of the secretion of digestive enzymes. The peptide hormone cholecystokinin (CCK) binds to CCK1R, which is an important pharmacological target for several diseases, including obesity. Interestingly, nutritional dietary peptides also appear to activate CCK1R, and may play a role in CCK1R signaling in the gut. In this study, a novel technique to screen for CCK1R ligands based on affinity-selection is described. Functional expressed CCK1R is reconstituted into membrane nanoparticles called NABBs (nanoscale apo-lipoprotein bound bilayers). NABBs are native-like bilayer membrane systems for incorporation of GPCRs. CCK1R-NABBs were characterized using a fluorescently labeled CCK analog and can be used as a cutting-edge technology to screen for CCK1R ligands using affinity-selection mass spectrometry.
authors.affiliation: Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium; Department of Food Safety and Food Quality, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.
authors.forename: Dorien
authors.initials: D
authors.lastname: Staljanssens
authors.affiliation: Laboratory of Chemical Biology & Signal Transduction, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Rico
authors.affiliation: Laboratory of Chemical Biology & Signal Transduction, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Minyoung
authors.initials: M
authors.lastname: Park
authors.affiliation: Department of Food Safety and Food Quality, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.
authors.forename: John
authors.initials: J
authors.lastname: Van Camp
authors.affiliation: Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.
authors.forename: Na
authors.initials: N
authors.lastname: Yu
authors.affiliation: Laboratory of Chemical Biology & Signal Transduction, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Huber
authors.affiliation: Laboratory of Chemical Biology & Signal Transduction, The Rockefeller University, New York, NY 10065, USA; Center for Alzheimer Research, Division for Neurogeriatrics, Department of Neurobiology, Care Sciences and Society, Karolinska Institute, SE-141 57 Huddinge, Sweden.
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Sakmar
authors.affiliation: Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium. Electronic address: guy.smagghe@ugent.be.
authors.forename: Guy
authors.initials: G
authors.lastname: Smagghe
mesh_terms: D000818:Animals; D001053:Apolipoproteins / Q000737:chemistry; D015374:Biosensing Techniques; D020013:Calcium Signaling; D004353:Drug Evaluation, Preclinical / Q000379:methods; D057809:HEK293 Cells; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry*; D021381:Protein Transport; D051381:Rats; D011949:Receptors, Cholecystokinin / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D029961:Zebrafish Proteins / Q000737:chemistry
chemical_list: D001053:Apolipoproteins; D011949:Receptors, Cholecystokinin; D029961:Zebrafish Proteins
keywords: Affinity-selection; Cholecystokinin; Cholecystokinin receptor type 1; GPCR; NABBs