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Biochim Biophys Acta

Title:		"In vitro characterization of ligand-induced oligomerization of the S. cerevisiae G-protein coupled receptor, Ste2p"
Author(s):		Shi C; Paige MF; Maley J; Loewen MC;
Address:		"Plant Biotechnology Institute, National Research Council of Canada, 110 Gymnasium Place, Saskatoon, SK, S7N 0W9, Canada"
Journal Title:		Biochim Biophys Acta
Year:		2009
Volume:		20081021
Issue:		1
Page Number:		1 - 7
DOI:		10.1016/j.bbagen.2008.10.003
ISSN/ISBN:		0006-3002 (Print) 0006-3002 (Linking)
Abstract:		"BACKGROUND: The S. cerevisiae alpha-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 alpha-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 alpha-factor induced stabilization of the dimeric form and higher order oligomeric forms of the receptor upon SDS-PAGE analysis. CONCLUSIONS: alpha-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"
Keywords:		"Biopolymers Cell Line Cell Membrane/chemistry Cross-Linking Reagents/chemistry Detergents Humans Ligands Light Maleimides/chemistry Mating Factor Micelles Microscopy, Atomic Force Peptides/chemistry Receptors, Mating Factor/agonists/*chemistry/genetics Re;"
Notes:		"MedlineShi, Chunhua Paige, Matthew F Maley, Jason Loewen, Michele C eng Research Support, Non-U.S. Gov't Netherlands 2008/11/11 Biochim Biophys Acta. 2009 Jan 1; 1790(1):1-7. doi: 10.1016/j.bbagen.2008.10.003. Epub 2008 Oct 21"