A Paradigm for Peptide Hormone-GPCR Analyses
Open Access
- 18 September 2020
- Vol. 25 (18), 4272
- https://doi.org/10.3390/molecules25184272
Abstract
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.Funding Information
- National Institutes of Health (GM22086, GM22087, GM 46520)
This publication has 179 references indexed in Scilit:
- Multiple regulatory roles of the carboxy terminus of Ste2p a yeast GPCRPharmacological Research, 2012
- Comparative NMR analysis of an 80-residue G protein-coupled receptor fragment in two membrane mimetic environmentsBiochimica et Biophysica Acta (BBA) - Biomembranes, 2011
- Changes in Conformation at the Cytoplasmic Ends of the Fifth and Sixth Transmembrane Helices of a Yeast G Protein-Coupled Receptor in Response to Ligand BindingBiochemistry, 2011
- Differential Interactions of Fluorescent Agonists and Antagonists with the Yeast G Protein Coupled Receptor Ste2pJournal of Molecular Biology, 2011
- Identification of Specific Transmembrane Residues and Ligand-Induced Interface Changes Involved In Homo-Dimer Formation of a Yeast G Protein-Coupled ReceptorBiochemistry, 2009
- Structure of a Double Transmembrane Fragment of a G-Protein-Coupled Receptor in MicellesBiophysical Journal, 2009
- Cross-Linking of a DOPA-Containing Peptide Ligand into Its G Protein-Coupled ReceptorBiochemistry, 2009
- G protein coupled receptor structure and activationBiochimica et Biophysica Acta (BBA) - Biomembranes, 2007
- A role for a complex between activated G protein-coupled receptors in yeast cellular matingProceedings of the National Academy of Sciences of the United States of America, 2007
- Double-Mutant Cycle Scanning of the Interaction of a Peptide Ligand and Its G Protein-Coupled ReceptorBiochemistry, 2007