Identification of a Cyanine-dye labeled peptidic ligand for Y₁R and Y₄R, based upon the Neuropeptide Y C-terminal analogue, BVD-15

Liu, Mengjie; Richardson, Rachel R.; Mountford, Simon J.; Zhang, Lei; Tempone, Matheus H.; Herzog, Herbert; Holliday, Nicholas D.; Thompson, Philip E.

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, [Lys²(sCy5), Arg⁴]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 [Lys²(sCy5), Arg⁴]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.

Citation

Liu, M., Richardson, R. R., Mountford, S. J., Zhang, L., Tempone, M. H., Herzog, H., Holliday, N. D., & Thompson, P. E. (2016). Identification of a Cyanine-dye labeled peptidic ligand for Y₁R and Y₄R, based upon the Neuropeptide Y C-terminal analogue, BVD-15. Bioconjugate Chemistry, 27(9), 2166-2175. https://doi.org/10.1021/acs.bioconjchem.6b00376

• Journal Article Type: Article
• Acceptance Date: Aug 8, 2016
• Online Publication Date: Aug 25, 2016
• Publication Date: Aug 11, 2016
• Deposit Date: Mar 23, 2017
• Publicly Available Date: Mar 23, 2017
• Journal: Bioconjugate Chemistry
• Print ISSN: 1043-1802
• Electronic ISSN: 1520-4812
• Publisher: American Chemical Society
• Peer Reviewed: Peer Reviewed
• Volume: 27
• Issue: 9
• Pages: 2166-2175
• DOI: https://doi.org/10.1021/acs.bioconjchem.6b00376
• Keywords: Neuropeptide Y, Fluorescent ligand, G protein coupled receptor, High content imaging
• Public URL: https://nottingham-repository.worktribe.com/output/805856
• Publisher URL: http://pubs.acs.org/doi/abs/10.1021/acs.bioconjchem.6b00376
• Additional Information: This document is the unedited author's version of a submitted work that was subsequently accepted for publication in the journal Bioconjugate Chemistry, Copyright © 2016 American Chemical Society after peer review. To access the final edited and published work see http://pubs.acs.org/doi/abs/10.1021/acs.bioconjchem.6b00376
• Contract Date: Mar 23, 2017