Kartierung von Protein-Peptid- und Protein-Proteinwechselwirkungen mittels genetisch kodierte Photocrosslinker
Zusammenfassung der Projektergebnisse
As major scientific achievements: we have revealed the mechanism of agonism and antagonism at the CRF1R; we have established a robust method to determine intermolecular pairs of proximal amino acids not only in ligand-receptor interfaces but in general in protein-protein interaction interfaces; we have developed a new system for the incorporation of Pyl-like ncAAs in mammalian cells and a general method for quantitative labeling of GPCRs on the surface of live cells. In this way, aims of project A and B of the original proposal are achieved. Although project C developed in a direction that was not planned at the beginning, it led to important results and nice publications. On these bases, projects involving 1) the study of ligand-GPCR interactions at other GPCRs, 2) the study of arrestin-GPCR interactions and 3) the generation of minimal sized FRET sensors for studies of receptor dynamics are ongoing and have a very promising long-term perspective.
Projektbezogene Publikationen (Auswahl)
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Incorporation of Unnatural Amino Acids into Proteins Expressed in Mammalian Cells. Methods Enzymol. 2016, 580, 89-107
Serfling, R.; Coin, I.
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Structural insight into the activation of a class B G-protein-coupled receptor by peptide hormones in live human cells. Elife 2017, 6
Seidel, L.; Zarzycka, B.; Zaidi, S. A.; Katritch, V.; Coin, I.
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Application of non-canonical crosslinking amino acids to study protein-protein interactions in live cells. Curr. Opin. Chem. Biol. 2018, 46, 156-163
Coin, I.
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Designer tRNAs for efficient incorporation of non-canonical amino acids by the pyrrolysine system in mammalian cells. Nucleic Acids Res. 2018, 46 (1), 1-10
Serfling, R.; Lorenz, C.; Etzel, M.; Schicht, G.; Böttke, T.; Mörl, M.; Coin, I.
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Mapping of Protein Interfaces in Live Cells Using Genetically Encoded Crosslinkers. Methods Mol. Biol. 2018, 1728, 221-235
Seidel, L.; Coin, I.
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Optimizing the genetic incorporation of chemical probes into GPCRs for photo-crosslinking mapping and bioorthogonal chemistry in live mammalian cells. J Vis Exp 2018, (134)
Serfling, R.; Seidel, L.; Bottke, T.; Coin, I.
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Exploring pairwise chemical crosslinking to study peptide-receptor interactions. ChemBioChem 2019, 20 (5), 683-692
Seidel, L.; Zarzycka, B.; Katritch, V.; Coin, I.
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Quantitative single-residue bioorthogonal labeling of G protein-coupled receptors in live cells. ACS Chem Biol 2019, 14 (6), 1141-1149
Serfling, R.; Seidel, L.; Bock, A.; Lohse, M. J.; Annibale, P.; Coin, I.
