A Paradigm for Peptide Hormone-GPCR Analyses
| Autor: | Jeffrey M. Becker, Fred Naider |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Peptide Hormones
Saccharomyces cerevisiae Mutant Pharmaceutical Science Peptide Review Ligands receptor-ligand interaction Analytical Chemistry Receptors G-Protein-Coupled lcsh:QD241-441 03 medical and health sciences photoactivated crosslinking Allosteric Regulation Protein Domains G protein-coupled receptors lcsh:Organic chemistry peptide analogs Drug Discovery Physical and Theoretical Chemistry Receptor 030304 developmental biology G protein-coupled receptor chemistry.chemical_classification 0303 health sciences Binding Sites biology 030302 biochemistry & molecular biology Organic Chemistry peptide pheromone Ligand (biochemistry) biology.organism_classification chemical crosslinking Transmembrane domain nuclear magnetic resonance chemistry Microscopy Fluorescence Chemistry (miscellaneous) receptor mutation Biophysics Molecular Medicine fluorescence screening Function (biology) Protein Binding |
| Zdroj: | Molecules, Vol 25, Iss 4272, p 4272 (2020) Molecules |
| ISSN: | 1420-3049 |
| Popis: | Work from our laboratories over the last 35 years that has focused on Ste2p, a G protein-coupled receptor (GPCR), and its tridecapeptide ligand α-factor is reviewed. Our work utilized the yeast Saccharomyces cerevisiae as a model system for understanding peptide-GPCR interactions. It explored the structure and function of synthetic α-factor analogs and biosynthetic receptor domains, as well as designed mutations of Ste2p. The results and conclusions are described using the nuclear magnetic resonance interrogation of synthetic Ste2p transmembrane domains (TMs), the fluorescence interrogation of agonist and antagonist binding, the biochemical crosslinking of peptide analogs to Ste2p, and the phenotypes of receptor mutants. We identified the ligand-binding domain in Ste2p, the functional assemblies of TMs, unexpected and interesting ligand analogs; gained insights into the bound α-factor structure; and unraveled the function and structures of various Ste2p domains, including the N-terminus, TMs, loops connecting the TMs, and the C-terminus. Our studies showed interactions between specific residues of Ste2p in an active state, but not resting state, and the effect of ligand activation on the dimerization of Ste2p. We show that, using a battery of different biochemical and genetic approaches, deep insight can be gained into the structure and conformational dynamics of GPCR-peptide interactions in the absence of a crystal structure. |
| Databáze: | OpenAIRE |
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