Project B04 – Mapping binding sites of NPY peptides on the Y5 receptor using genetically encoded crosslinkers

Very little is known about ligand binding to the Y5 receptor (Y5R) and its activation. We will use genetically encoded crosslinkers to map on the Y5R the binding paths of its three natural peptide ligands and identify inter-molecular pairs of vicinal amino acids. Spatial constraints will be derived, which will serve to build accurate computational models of the peptide-Y5R complexes. Analysing the models in the frame of existing data for other Y receptors will help identifying discriminants for selectivity and shed light on the evolution of the Y system.

Contact

Resources

We focus on mapping protein-protein interaction surfaces in the live cell by combining chemical tools and modern molecular biology techniques. In particular, we use the expanded genetic code technology to incorporate crosslinking amino acids into proteins at specific sites.

Publications

Coming soon.

Coin I, Katritch V, Sun T, Xiang Z, Siu FY, Beyermann M, Stevens RC, Wang L. Genetically encoded chemical probes reveal the binding path of Urocortin-I to CRF class B GPCR. 2013; 155:1258-69.

Seidel L, Zarzycka B, Zaidi SA, Katritch V, Coin I. Structural insight into the activation of a class B G-protein-coupled receptor by peptide hormones in live human cells. 2017; 6:27711.

Coin I, Perrin MH, Vale WW, Wang L. Photocrosslinkers incorporated into G-Protein-Coupled-Receptors in mammalian cells: a ligand comparison. Selected as Very-Important-Paper (VIP) by referees. Angew Chem Int Ed Engl. 2011; 50:8077-81.

Serfling R, Lorenz C, Etzel M, Schicht G, Böttke T, Mörl M, Coin I. Designer tRNAs for efficient incorporation of non-canonical amino acids by the pyrrolysine system in mammalian cells. Selected as breakthrough paper, journal cover. Nucleic Acids Res. 2018; 46:1-10.

Serfling R, Seidel L, Böttke T, Coin I. Optimizing the genetic incorporation of chemical probes into GPCRs for photo-crosslinking mapping and bioorthogonal chemistry in live mammalian cells. J Vis 2018; 134.

Xiang Z, Ren H, Hu YS, Coin I, Wei J, Cang H, Wang L. Adding an unnatural covalent bond to proteins through proximity-enhanced bioreactivity. Nat Methods. 2013; 10:885-8.

Coin I. Application of non-canonical crosslinking amino acids to study protein-protein interactions in live cells. Curr Opin Chem biol. 2018; 46:156-63.

Seidel L, Zarzycka B, Katritch V, Coin I. Exploring pairwise chemical crosslinking to study peptide-receptor interactions. ChemBioChem, 2019; 20:1-11.

Coin I, Beyermann M, Bienert M. Solid-phase peptide synthesis: from standard procedures to the synthesis of difficult sequences. Nat Protoc. 2007; 2:3247-56.