title: Identification of a Cyanine-Dye Labeled Peptidic Ligand for Y1R and Y4R, Based upon the Neuropeptide Y C-Terminal Analogue, BVD-15.
abstract: Traceable truncated Neuropeptide Y (NPY) analogues with Y1 receptor (Y1R) affinity and selectivity are highly desirable tools in studying receptor location, regulation, and biological functions. A range of fluorescently labeled analogues of a reported Y1R/Y4R preferring ligand BVD-15 have been prepared and evaluated using high content imaging techniques. One peptide, [Lys(2)(sCy5), Arg(4)]BVD-15, was characterized as an Y1R antagonist with a pKD of 7.2 measured by saturation analysis using fluorescent imaging. The peptide showed 8-fold lower affinity for Y4R (pKD = 6.2) and was a partial agonist at this receptor. The suitability of [Lys(2)(sCy5), Arg(4)]BVD-15 for Y1R and Y4R competition binding experiments was also demonstrated in intact cells. The nature of the label was shown to be critical with replacement of sCy5 by the more hydrophobic Cy5.5 resulting in a switch from Y1R antagonist to Y1R partial agonist.
authors: Medicinal Chemistry, Monash Institute of Pharmaceutical Sciences, Monash University , 381 Royal Parade, Parkville, VIC 3052, Australia.
authors: Mengjie
authors: M
authors: Liu
authors: Cell Signalling Research Group, School of Life Sciences, University of Nottingham, Queen's Medical Centre , Nottingham, NG7 2UH, United Kingdom.
authors: Rachel R
authors: RR
authors: Richardson
authors: Medicinal Chemistry, Monash Institute of Pharmaceutical Sciences, Monash University , 381 Royal Parade, Parkville, VIC 3052, Australia.
authors: Simon J
authors: SJ
authors: Mountford
authors: Neuroscience Division, Garvan Institute of Medical Research, St. Vincent's Hospital , Darlinghurst, NSW 2010, Australia.
authors: Lei
authors: L
authors: Zhang
authors: Cell Signalling Research Group, School of Life Sciences, University of Nottingham, Queen's Medical Centre , Nottingham, NG7 2UH, United Kingdom.
authors: Matheus H
authors: MH
authors: Tempone
authors: Neuroscience Division, Garvan Institute of Medical Research, St. Vincent's Hospital , Darlinghurst, NSW 2010, Australia.
authors: Herbert
authors: H
authors: Herzog
authors: Cell Signalling Research Group, School of Life Sciences, University of Nottingham, Queen's Medical Centre , Nottingham, NG7 2UH, United Kingdom.
authors: Nicholas D
authors: ND
authors: Holliday
authors: Medicinal Chemistry, Monash Institute of Pharmaceutical Sciences, Monash University , 381 Royal Parade, Parkville, VIC 3052, Australia.
authors: Philip E
authors: PE
authors: Thompson
mesh_terms: D000595:Amino Acid Sequence; D001667:Binding, Competitive; D002232:Carbocyanines / Q000737:chemistry*; D004396:Coloring Agents / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D008024:Ligands; D009478:Neuropeptide Y / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism*; D017476:Receptors, Neuropeptide Y / Q000378:metabolism*; D013194:Staining and Labeling
chemical_list: D002232:Carbocyanines; D004396:Coloring Agents; D008024:Ligands; D009478:Neuropeptide Y; D010446:Peptide Fragments; D017476:Receptors, Neuropeptide Y