NPY receptors
Obesity[1], neurodegenerative diseases, stress, anxiety disorders[2] and addictive behaviour[3] are the outcome of complex signalling mechanisms in the mammalian central nervous system, crucially influenced by Neuropeptide Y (NPY) and its pharmacological targets, the NPY family of G-protein coupled receptors. Represented by four different receptor subtypes, the NPY receptors are frequently discussed pharmacological targets, in particular the NPY Y1 receptor subtype, which our working group is focusing on. Mediated by a Gi/o-coupled intracellular signalling pathway, binding of the endogenous agonist NPY to the NPY Y1 receptor results in increased food intake, increased blood pressure and anxiolytic effects.[4] In the last decade, the overexpression of NPY receptor subtypes in various cancer tissues has also been reported, further complementing the medicinal relevance of this receptor family.[5]
We are in particular investigating on receptor-ligand interactions utilizing fluorescence anisotropy (FA) based assays, which have been already successfully applied on various other GPCRs in this working group.[6,7] Therefore, we use NPY Y1R-expressing baculovirus particles as receptor sources instead of live cells. Supplied with highly affine and selective fluorescent NPY Y1 tracers by our collaboration partners around Dr. Max Keller from the University of Regensburg , we emphasize on the analysis of binding kinetics, the investigation on potential allosteric modulation of receptor binding and the development of fluorescence-based competition binding assays as an alternative to radiometric methods. So far, the FA assay was successfully applied to characterize the receptor binding of both fluorescent NPY Y1 agonists and antagonists. We also have the know-how to study the functional behaviour of NPY ligands by using genetically encoded Epac-based FRET biosensor BacMam system.
[1] Hofmann, S.; Bellmann-Sickert, K.; Beck-Sickinger Annette, G. In Biological Chemistry 2019; Vol. 400, p 299.
[2] Reichmann, F.; Holzer, P. Neuropeptides 2016, 55, 99-109.
[3] Thorsell, A.; Mathé, A. A. Frontiers in Endocrinology 2017, 8.
[4] British Journal of Pharmacology 2009, 158, S74-S74.
[5] Li, J.; Tian, Y.; Wu, A. Regenerative Biomaterials 2015, 2, 215-219.
[6] Link, R.; Kopanchuk, S.; Rinken, A. SpringerPlus 2015, 4, P23.
[7] Allikalt, A.; Rinken, A. SpringerPlus 2015, 4.
We are in particular investigating on receptor-ligand interactions utilizing fluorescence anisotropy (FA) based assays, which have been already successfully applied on various other GPCRs in this working group.[6,7] Therefore, we use NPY Y1R-expressing baculovirus particles as receptor sources instead of live cells. Supplied with highly affine and selective fluorescent NPY Y1 tracers by our collaboration partners around Dr. Max Keller from the University of Regensburg , we emphasize on the analysis of binding kinetics, the investigation on potential allosteric modulation of receptor binding and the development of fluorescence-based competition binding assays as an alternative to radiometric methods. So far, the FA assay was successfully applied to characterize the receptor binding of both fluorescent NPY Y1 agonists and antagonists. We also have the know-how to study the functional behaviour of NPY ligands by using genetically encoded Epac-based FRET biosensor BacMam system.
[1] Hofmann, S.; Bellmann-Sickert, K.; Beck-Sickinger Annette, G. In Biological Chemistry 2019; Vol. 400, p 299.
[2] Reichmann, F.; Holzer, P. Neuropeptides 2016, 55, 99-109.
[3] Thorsell, A.; Mathé, A. A. Frontiers in Endocrinology 2017, 8.
[4] British Journal of Pharmacology 2009, 158, S74-S74.
[5] Li, J.; Tian, Y.; Wu, A. Regenerative Biomaterials 2015, 2, 215-219.
[6] Link, R.; Kopanchuk, S.; Rinken, A. SpringerPlus 2015, 4, P23.
[7] Allikalt, A.; Rinken, A. SpringerPlus 2015, 4.