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Peptides. 2015 Jun;68:219-27. doi: 10.1016/j.peptides.2014.10.015. Epub 2014 Nov 5.

Development of a CCK1R-membrane nanoparticle as a fish-out tool for bioactive peptides.

Author information

1
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.
2
Laboratory of Chemical Biology & Signal Transduction, The Rockefeller University, New York, NY 10065, USA.
3
Department of Food Safety and Food Quality, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.
4
Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.
5
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.
6
Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium. Electronic address: guy.smagghe@ugent.be.

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.

KEYWORDS:

Affinity-selection; Cholecystokinin; Cholecystokinin receptor type 1; GPCR; NABBs

PMID:
25451329
DOI:
10.1016/j.peptides.2014.10.015
[Indexed for MEDLINE]

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