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J Phys Chem B

Title:		Ste2p Under the Microscope: the Investigation of Oligomeric States of a Yeast G Protein-Coupled Receptor
Author(s):		Cevheroglu O; Murat M; Mingu-Akmete S; Son CD;
Address:		"Stem Cell Institute, Ankara University, Cankaya, 06520 Ankara, Turkey. Department of Biological Sciences, Middle East Technical University, Cankaya, 06800 Ankara, Turkey"
Journal Title:		J Phys Chem B
Year:		2021
Volume:		20210817
Issue:		33
Page Number:		9526 - 9536
DOI:		10.1021/acs.jpcb.1c05872
ISSN/ISBN:		1520-5207 (Electronic) 1520-5207 (Linking)
Abstract:		"Oligomerization of G protein-coupled receptors (GPCRs) may play important roles in maturation, internalization, signaling, and pharmacology of these receptors. However, the nature and extent of their oligomerization is still under debate. In our study, Ste2p, a yeast mating pheromone GPCR, was tagged with enhanced green fluorescent protein (EGFP), mCherry, and with split florescent protein fragments at the receptor C-terminus. The Forster resonance energy transfer (FRET) technique was used to detect receptors' oligomerization by calculating the energy transfer from EGFP to mCherry. Stimulation of Ste2p oligomers with the receptor ligand did not result in any significant change on observed FRET values. The bimolecular fluorescence complementation (BiFC) assay was combined with FRET to further investigate the tetrameric complexes of Ste2p. Our results suggest that in its quiescent (nonligand-activated) state, Ste2p is found at least as a tetrameric complex on the plasma membrane. Intriguingly, receptor tetramers in their active form showed a significant increase in FRET. This study provides a direct in vivo visualization of Ste2p tetramers and the pheromone effect on the extent of the receptor oligomerization"
Keywords:		"Fluorescence Resonance Energy Transfer Protein Binding Receptors, G-Protein-Coupled/genetics/metabolism Receptors, Mating Factor/genetics/metabolism *Saccharomyces cerevisiae/genetics *Saccharomyces cerevisiae Proteins/genetics/metabolism;"
Notes:		"MedlineCevheroglu, Orkun Murat, Merve Mingu-Akmete, Sara Son, Cagdas D eng Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't 2021/08/27 J Phys Chem B. 2021 Aug 26; 125(33):9526-9536. doi: 10.1021/acs.jpcb.1c05872. Epub 2021 Aug 17"