In vitro characterization of ligand-induced oligomerization of the S. cerevisiae G-protein coupled receptor, Ste2p

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DOIResolve DOI: http://doi.org/10.1016/j.bbagen.2008.10.003
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TypeArticle
Journal titleBiochimica et Biophysica Acta (BBA) - General Subjects
Volume1790
Issue1
Pages17; # of pages: 7
SubjectG-protein coupled receptor; Ste2p; Ligand-induced oligomerization; Chemical crosslinking; Dynamic light scattering; Atomic force microscopy
AbstractBackground: The S. cerevisiae α-factor receptor, Ste2p, is a G-protein coupled receptor that plays key roles in yeast signaling and mating. Oligomerization of Ste2p has previously been shown to be important for intracellular trafficking, receptor processing and endocytosis. However the role of ligand in receptor oligomerization remains enigmatic. Methods: Using functional recombinant forms of purified Ste2p, atomic force microscopy, dynamic light scattering and chemical crosslinking are applied to investigate the role of ligand in Ste2p oligomerization. Results: Atomic force microscopy images indicate a molecular height for recombinant Ste2p in the presence of α-factor nearly double that of Ste2p alone. This observation is supported by complementary dynamic light scattering measurements which indicate a ligand-induced increase in the polydispersity of the Ste2p hydrodynamic radius. Finally, chemical cross-linking of HEK293 plasma membranes presenting recombinant Ste2p indicates α-factor induced stabilization of the dimeric form and higher order oligomeric forms of the receptor upon SDS-PAGE analysis. Conclusions: α-factor induces oligomerization of Ste2p in vitro and in membrane. General significance: These results provide additional evidence of a possible role for ligand in mediation of Ste2p oligomerization in vivo.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada; NRC Plant Biotechnology Institute
Peer reviewedNo
NPARC number21187431
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Record identifier3b64308e-c0e8-47fb-a7ff-54ddeefe18aa
Record created2013-01-10
Record modified2016-05-09
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