Development of a CCK1R-membrane nanoparticle as a fish-out tool for bioactive peptides |
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| Affiliation: | 1. Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium;2. Department of Food Safety and Food Quality, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium;3. Laboratory of Chemical Biology & Signal Transduction, The Rockefeller University, New York, NY 10065, USA;4. Center for Alzheimer Research, Division for Neurogeriatrics, Department of Neurobiology, Care Sciences and Society, Karolinska Institute, SE-141 57 Huddinge, Sweden;1. Centro de Investigaciones Sobre Enfermedades Infecciosas, Instituto Nacional de Salud Pública, Av. Universidad 655, Cuernavaca, Morelos 62100, Mexico;2. Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Postal 510-3, Cuernavaca 62250, Morelos, Mexico;1. Department of Applied Chemistry, College of Science, China Agricultural University, Beijing 100193, PR China;2. Department of Cell and Systems Biology, University of Toronto, 25 Harbord St., Toronto, ON, Canada M5S 3G5;3. State Key Laboratory of the Discovery and Development of Novel Pesticide, Shenyang Research Institute of Chemical Industry Co. Ltd., Shenyang 110021, PR China;1. Department of Protozoology, Faculty of Tropical Medicine, Mahidol University Ratchathewi, Bangkok 10400, Thailand;2. Bacterial Protein Toxin Research Group, Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Nakornpathom 73170, Thailand |
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| 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. |
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| Keywords: | Cholecystokinin receptor type 1 Cholecystokinin NABBs GPCR Affinity-selection |
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