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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
4
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pubmed:dateCreated |
2011-5-30
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pubmed:abstractText |
We describe a rapid method to probe for mutations in cell surface ligand-binding proteins that affect the environment of bound ligand. The method uses fluorescence-activated cell sorting to screen randomly mutated receptors for substitutions that alter the fluorescence emission spectrum of environmentally sensitive fluorescent ligands. When applied to the yeast ?-factor receptor Ste2p, a G protein-coupled receptor, the procedure identified 22 substitutions that red shift the emission of a fluorescent agonist, including substitutions at residues previously implicated in ligand binding and at additional sites. A separate set of substitutions, identified in a screen for mutations that alter the emission of a fluorescent ?-factor antagonist, occurs at sites that are unlikely to contact the ligand directly. Instead, these mutations alter receptor conformation to increase ligand-binding affinity and provide signaling in response to antagonists of normal receptors. These results suggest that receptor--agonist interactions involve at least two sites, of which only one is specific for the activated conformation of the receptor.
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pubmed:grant |
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jun
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pubmed:issn |
1089-8638
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pubmed:author |
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pubmed:copyrightInfo |
Copyright © 2011. Published by Elsevier Ltd.
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pubmed:issnType |
Electronic
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pubmed:day |
17
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pubmed:volume |
409
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
513-28
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pubmed:dateRevised |
2011-9-26
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pubmed:meshHeading |
pubmed-meshheading:21477594-Amino Acid Sequence,
pubmed-meshheading:21477594-Flow Cytometry,
pubmed-meshheading:21477594-Fluorescent Dyes,
pubmed-meshheading:21477594-Ligands,
pubmed-meshheading:21477594-Molecular Sequence Data,
pubmed-meshheading:21477594-Mutation,
pubmed-meshheading:21477594-Protein Binding,
pubmed-meshheading:21477594-Protein Conformation,
pubmed-meshheading:21477594-Receptors, Mating Factor,
pubmed-meshheading:21477594-Saccharomyces cerevisiae,
pubmed-meshheading:21477594-Saccharomyces cerevisiae Proteins
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pubmed:year |
2011
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pubmed:articleTitle |
Differential interactions of fluorescent agonists and antagonists with the yeast G protein coupled receptor Ste2p.
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pubmed:affiliation |
Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, NY 14642, USA.
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pubmed:publicationType |
Journal Article,
Research Support, N.I.H., Extramural
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