The yeast Ste2p G protein-coupled receptor dimerizes on the cell plasma membrane

Date

2017-05-01

Author

Cevheroglu, Orkun
Kumas, Gozde
Hauser, Melinda
Becker, Jeffrey M.
Son, Çağdaş Devrim

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Dimerization of G protein-coupled receptors (GPCR) may play an important role in maturation, internalization, signaling and/or pharmacology of these receptors. However, the location where dimerization occurs is still under debate. In our study, variants of Ste2p, a yeast mating pheromone GPCR, were tagged with split EGFP (enhanced green fluorescent protein) fragments inserted between transmembrane domain seven and the C-terminus or appended to the C-terminus. Bimolecular Fluorescence Complementation (BiFC) assay was used to determine where receptor dimerization occurred during protein trafficking by monitoring generation of EGFP fluorescence, which occurred upon GPCR dimerization. Our results suggest that these tagged receptors traffic to the membrane as monomers, undergo dimerization or higher ordered oligomerization predominantly on the plasma membrane, and are internalized as dimers/oligomers. This study is the first to provide direct in vivo visualization of GPCR dimerization/oligomerization, during trafficking to and from the plasma membrane.

Subject Keywords

Ste2p, Split EGFP, Bimolecular fluorescence complementation assay (BiFC), GPCR traffic, GPCR dimerization

URI

https://hdl.handle.net/11511/41182

Journal

BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES

DOI

https://doi.org/10.1016/j.bbamem.2017.01.008

Collections

Department of Biology, Article

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Citation Formats

O. Cevheroglu, G. Kumas, M. Hauser, J. M. Becker, and Ç. D. Son, “The yeast Ste2p G protein-coupled receptor dimerizes on the cell plasma membrane,” BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, pp. 698–711, 2017, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/41182.