| http://purl.uniprot.org/citations/20923758 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/20923758 | http://www.w3.org/2000/01/rdf-schema#comment | "Fundamental knowledge about how G protein-coupled receptors and their ligands interact is important for understanding receptor-ligand binding and the development of new drug discovery strategies. We have used cross-linking and tandem mass spectrometry analyses to investigate the interaction of the N terminus of the Saccharomyces cerevisiae tridecapeptide pheromone, α-factor (WHWLQLKPGQPMY), and Ste2p, its cognate G protein-coupled receptor. The Trp(1) residue of α-factor was replaced by 3,4-dihydroxyphenylalanine (DOPA) for periodate-mediated chemical cross-linking, and biotin was conjugated to Lys(7) for detection purposes to create the peptide [DOPA(1),Lys(7)(BioACA),Nle(12)]α-factor, called Bio-DOPA(1)-α-factor. This ligand analog was a potent agonist and bound to Ste2p with ∼65 nanomolar affinity. Immunoblot analysis of purified Ste2p samples that were treated with Bio-DOPA(1)-α-factor showed that the peptide analog cross-linked efficiently to Ste2p. The cross-linking was inhibited by the presence of either native α-factor or an α-factor antagonist. MALDI-TOF and immunoblot analyses revealed that Bio-DOPA(1)-α-factor cross-linked to a fragment of Ste2p encompassing residues Ser(251)-Met(294). Fragmentation of the cross-linked fragment and Ste2p using tandem mass spectrometry pinpointed the cross-link point of the DOPA(1) of the α-factor analog to the Ste2p Lys(269) side chain near the extracellular surface of the TM6-TM7 bundle. This conclusion was confirmed by a greatly diminished cross-linking of Bio-DOPA(1)-α-factor into a Ste2p(K269A) mutant. Based on these and previously obtained binding contact data, a mechanism of α-factor binding to Ste2p is proposed. The model for bound α-factor shows how ligand binding leads to conformational changes resulting in receptor activation of the signal transduction pathway."xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://purl.org/dc/terms/identifier | "doi:10.1074/jbc.m110.149500"xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://purl.uniprot.org/core/author | "Huang L."xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://purl.uniprot.org/core/author | "Link A.J."xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://purl.uniprot.org/core/author | "Becker J.M."xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://purl.uniprot.org/core/author | "Umanah G.K."xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://purl.uniprot.org/core/author | "Ding F.X."xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://purl.uniprot.org/core/author | "Arshava B."xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://purl.uniprot.org/core/author | "Naider F."xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://purl.uniprot.org/core/author | "Farley A.R."xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://purl.uniprot.org/core/date | "2010"xsd:gYear |
| http://purl.uniprot.org/citations/20923758 | http://purl.uniprot.org/core/name | "J Biol Chem"xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://purl.uniprot.org/core/pages | "39425-39436"xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://purl.uniprot.org/core/title | "Identification of residue-to-residue contact between a peptide ligand and its G protein-coupled receptor using periodate-mediated dihydroxyphenylalanine cross-linking and mass spectrometry."xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://purl.uniprot.org/core/volume | "285"xsd:string |
| http://purl.uniprot.org/citations/20923758 | http://www.w3.org/2004/02/skos/core#exactMatch | http://purl.uniprot.org/pubmed/20923758 |
| http://purl.uniprot.org/citations/20923758 | http://xmlns.com/foaf/0.1/primaryTopicOf | https://pubmed.ncbi.nlm.nih.gov/20923758 |
| http://purl.uniprot.org/uniprot/#_D6VTK4-mappedCitation-20923758 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/20923758 |
| http://purl.uniprot.org/uniprot/#_P01149-mappedCitation-20923758 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/20923758 |
| http://purl.uniprot.org/uniprot/#_P32435-mappedCitation-20923758 | http://www.w3.org/1999/02/22-rdf-syntax-ns#object | http://purl.uniprot.org/citations/20923758 |
| http://purl.uniprot.org/uniprot/D6VTK4 | http://purl.uniprot.org/core/mappedCitation | http://purl.uniprot.org/citations/20923758 |
| http://purl.uniprot.org/uniprot/P32435 | http://purl.uniprot.org/core/mappedCitation | http://purl.uniprot.org/citations/20923758 |
| http://purl.uniprot.org/uniprot/P01149 | http://purl.uniprot.org/core/mappedCitation | http://purl.uniprot.org/citations/20923758 |