Detecting ligand interactions with G protein-coupled receptors in real-time on living cells.
Identifieur interne : 000797 ( Main/Exploration ); précédent : 000796; suivant : 000798Detecting ligand interactions with G protein-coupled receptors in real-time on living cells.
Auteurs : RBID : pubmed:24211197English descriptors
- KwdEn :
- Animals, Bombesin (analogs & derivatives), HEK293 Cells, Humans, Ligands, Peptide YY (chemistry), Peptide YY (metabolism), Radioactive Tracers, Radioligand Assay, Receptors, G-Protein-Coupled (metabolism), Receptors, Neuropeptide Y (genetics), Receptors, Neuropeptide Y (metabolism), Swine, Time Factors, Transfection.
- MESH :
- chemical , analogs & derivatives : Bombesin.
- chemical , chemistry : Peptide YY.
- chemical , genetics : Receptors, Neuropeptide Y.
- chemical , metabolism : Peptide YY, Receptors, G-Protein-Coupled, Receptors, Neuropeptide Y.
- Animals, HEK293 Cells, Humans, Ligands, Radioactive Tracers, Radioligand Assay, Swine, Time Factors, Transfection.
Abstract
G protein-coupled receptors (GPCRs) are a large group of receptors of great biological and clinical relevance. Despite this, the tools for a detailed analysis of ligand-GPCR interactions are limited. The aim of this paper was to demonstrate how ligand binding to GPCRs can be followed in real-time on living cells. This was conducted using two model systems, the radiolabeled porcine peptide YY (pPYY) interacting with transfected human Y2 receptor (hY2R) and the bombesin antagonist RM26 binding to the naturally expressed gastrin-releasing peptide receptor (GRPR). By following the interaction over time, the affinity and kinetic properties such as association and dissociation rate were obtained. Additionally, data were analyzed using the Interaction Map method, which can evaluate a real-time binding curve and present the number of parallel interactions contributing to the curve. It was found that pPYY binds very slowly with an estimated time to equilibrium of approximately 12h. This may be problematic in standard end-point assays where equilibrium is required. The RM26 binding showed signs of heterogeneity, observed as two parallel interactions with unique kinetic properties. In conclusion, measuring binding in real-time using living cells opens up for a better understanding of ligand interactions with GPCRs.
DOI: 10.1016/j.bbrc.2013.10.149
PubMed: 24211197
Links toward previous steps (curation, corpus...)
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Detecting ligand interactions with G protein-coupled receptors in real-time on living cells.</title><author><name sortKey="Xu, Bo" uniqKey="Xu B">Bo Xu</name><affiliation wicri:level="1"><nlm:affiliation>Department of Neuroscience, Unit of Pharmacology, Uppsala University, Husargatan 2, 751 24 Uppsala, Sweden. Electronic address: bo.xu@neuro.uu.se.</nlm:affiliation><country xml:lang="fr">Suède</country><wicri:regionArea>Department of Neuroscience, Unit of Pharmacology, Uppsala University, Husargatan 2, 751 24 Uppsala</wicri:regionArea></affiliation></author><author><name sortKey="Varasteh, Zohreh" uniqKey="Varasteh Z">Zohreh Varasteh</name></author><author><name sortKey="Orlova, Anna" uniqKey="Orlova A">Anna Orlova</name></author><author><name sortKey="Andersson, Karl" uniqKey="Andersson K">Karl Andersson</name></author><author><name sortKey="Larhammar, Dan" uniqKey="Larhammar D">Dan Larhammar</name></author><author><name sortKey="Bj Rkelund, Hanna" uniqKey="Bj Rkelund H">Hanna Björkelund</name></author></titleStmt><publicationStmt><date when="2013">2013</date><idno type="RBID">pubmed:24211197</idno><idno type="pmid">24211197</idno><idno type="doi">10.1016/j.bbrc.2013.10.149</idno><idno type="wicri:Area/Main/Corpus">000316</idno><idno type="wicri:Area/Main/Curation">000316</idno><idno type="wicri:Area/Main/Exploration">000797</idno></publicationStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Bombesin (analogs & derivatives)</term><term>HEK293 Cells</term><term>Humans</term><term>Ligands</term><term>Peptide YY (chemistry)</term><term>Peptide YY (metabolism)</term><term>Radioactive Tracers</term><term>Radioligand Assay</term><term>Receptors, G-Protein-Coupled (metabolism)</term><term>Receptors, Neuropeptide Y (genetics)</term><term>Receptors, Neuropeptide Y (metabolism)</term><term>Swine</term><term>Time Factors</term><term>Transfection</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analogs & derivatives" xml:lang="en"><term>Bombesin</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Peptide YY</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Receptors, Neuropeptide Y</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Peptide YY</term><term>Receptors, G-Protein-Coupled</term><term>Receptors, Neuropeptide Y</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>HEK293 Cells</term><term>Humans</term><term>Ligands</term><term>Radioactive Tracers</term><term>Radioligand Assay</term><term>Swine</term><term>Time Factors</term><term>Transfection</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">G protein-coupled receptors (GPCRs) are a large group of receptors of great biological and clinical relevance. Despite this, the tools for a detailed analysis of ligand-GPCR interactions are limited. The aim of this paper was to demonstrate how ligand binding to GPCRs can be followed in real-time on living cells. This was conducted using two model systems, the radiolabeled porcine peptide YY (pPYY) interacting with transfected human Y2 receptor (hY2R) and the bombesin antagonist RM26 binding to the naturally expressed gastrin-releasing peptide receptor (GRPR). By following the interaction over time, the affinity and kinetic properties such as association and dissociation rate were obtained. Additionally, data were analyzed using the Interaction Map method, which can evaluate a real-time binding curve and present the number of parallel interactions contributing to the curve. It was found that pPYY binds very slowly with an estimated time to equilibrium of approximately 12h. This may be problematic in standard end-point assays where equilibrium is required. The RM26 binding showed signs of heterogeneity, observed as two parallel interactions with unique kinetic properties. In conclusion, measuring binding in real-time using living cells opens up for a better understanding of ligand interactions with GPCRs.</div></front></TEI><pubmed><MedlineCitation Owner="NLM" Status="MEDLINE"><PMID Version="1">24211197</PMID><DateCreated><Year>2013</Year><Month>12</Month><Day>02</Day></DateCreated><DateCompleted><Year>2014</Year><Month>03</Month><Day>31</Day></DateCompleted><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1090-2104</ISSN><JournalIssue CitedMedium="Internet"><Volume>441</Volume><Issue>4</Issue><PubDate><Year>2013</Year><Month>Nov</Month><Day>29</Day></PubDate></JournalIssue><Title>Biochemical and biophysical research communications</Title><ISOAbbreviation>Biochem. Biophys. Res. Commun.</ISOAbbreviation></Journal><ArticleTitle>Detecting ligand interactions with G protein-coupled receptors in real-time on living cells.</ArticleTitle><Pagination><MedlinePgn>820-4</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.bbrc.2013.10.149</ELocationID><ELocationID EIdType="pii" ValidYN="Y">S0006-291X(13)01842-1</ELocationID><Abstract><AbstractText>G protein-coupled receptors (GPCRs) are a large group of receptors of great biological and clinical relevance. Despite this, the tools for a detailed analysis of ligand-GPCR interactions are limited. The aim of this paper was to demonstrate how ligand binding to GPCRs can be followed in real-time on living cells. This was conducted using two model systems, the radiolabeled porcine peptide YY (pPYY) interacting with transfected human Y2 receptor (hY2R) and the bombesin antagonist RM26 binding to the naturally expressed gastrin-releasing peptide receptor (GRPR). By following the interaction over time, the affinity and kinetic properties such as association and dissociation rate were obtained. Additionally, data were analyzed using the Interaction Map method, which can evaluate a real-time binding curve and present the number of parallel interactions contributing to the curve. It was found that pPYY binds very slowly with an estimated time to equilibrium of approximately 12h. This may be problematic in standard end-point assays where equilibrium is required. The RM26 binding showed signs of heterogeneity, observed as two parallel interactions with unique kinetic properties. In conclusion, measuring binding in real-time using living cells opens up for a better understanding of ligand interactions with GPCRs.</AbstractText><CopyrightInformation>Copyright © 2013 Elsevier Inc. All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Xu</LastName><ForeName>Bo</ForeName><Initials>B</Initials><Affiliation>Department of Neuroscience, Unit of Pharmacology, Uppsala University, Husargatan 2, 751 24 Uppsala, Sweden. Electronic address: bo.xu@neuro.uu.se.</Affiliation></Author><Author ValidYN="Y"><LastName>Varasteh</LastName><ForeName>Zohreh</ForeName><Initials>Z</Initials></Author><Author ValidYN="Y"><LastName>Orlova</LastName><ForeName>Anna</ForeName><Initials>A</Initials></Author><Author ValidYN="Y"><LastName>Andersson</LastName><ForeName>Karl</ForeName><Initials>K</Initials></Author><Author ValidYN="Y"><LastName>Larhammar</LastName><ForeName>Dan</ForeName><Initials>D</Initials></Author><Author ValidYN="Y"><LastName>Björkelund</LastName><ForeName>Hanna</ForeName><Initials>H</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType>Journal Article</PublicationType><PublicationType>Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2013</Year><Month>11</Month><Day>06</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Biochem Biophys Res Commun</MedlineTA><NlmUniqueID>0372516</NlmUniqueID><ISSNLinking>0006-291X</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance>Ligands</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance>Radioactive Tracers</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance>Receptors, G-Protein-Coupled</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance>Receptors, Neuropeptide Y</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance>neuropeptide Y2 receptor</NameOfSubstance></Chemical><Chemical><RegistryNumber>106388-42-5</RegistryNumber><NameOfSubstance>Peptide YY</NameOfSubstance></Chemical><Chemical><RegistryNumber>PX9AZU7QPK</RegistryNumber><NameOfSubstance>Bombesin</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Bombesin</DescriptorName><QualifierName MajorTopicYN="N">analogs & derivatives</QualifierName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">HEK293 Cells</DescriptorName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Ligands</DescriptorName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Peptide YY</DescriptorName><QualifierName MajorTopicYN="N">chemistry</QualifierName><QualifierName MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Radioactive Tracers</DescriptorName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="Y">Radioligand Assay</DescriptorName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Receptors, G-Protein-Coupled</DescriptorName><QualifierName MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Receptors, Neuropeptide Y</DescriptorName><QualifierName MajorTopicYN="N">genetics</QualifierName><QualifierName MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Swine</DescriptorName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Time Factors</DescriptorName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Transfection</DescriptorName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">(111)In</Keyword><Keyword MajorTopicYN="N">(125)I</Keyword><Keyword MajorTopicYN="N">G protein-coupled receptor</Keyword><Keyword MajorTopicYN="N">GPCR</Keyword><Keyword MajorTopicYN="N">GRPR</Keyword><Keyword MajorTopicYN="N">Heterogeneity</Keyword><Keyword MajorTopicYN="N">Interaction Map</Keyword><Keyword MajorTopicYN="N">Kinetics</Keyword><Keyword MajorTopicYN="N">LigandTracer</Keyword><Keyword MajorTopicYN="N">Real-time</Keyword><Keyword MajorTopicYN="N">gastrin-releasing peptide receptor</Keyword><Keyword MajorTopicYN="N">hY2R</Keyword><Keyword MajorTopicYN="N">human Y2 receptor</Keyword><Keyword MajorTopicYN="N">indium-111</Keyword><Keyword MajorTopicYN="N">iodine-125</Keyword><Keyword MajorTopicYN="N">pPYY</Keyword><Keyword MajorTopicYN="N">porcine peptide YY</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2013</Year><Month>10</Month><Day>23</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2013</Year><Month>10</Month><Day>28</Day></PubMedPubDate><PubMedPubDate PubStatus="aheadofprint"><Year>2013</Year><Month>11</Month><Day>6</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2013</Year><Month>11</Month><Day>12</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2013</Year><Month>11</Month><Day>12</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2014</Year><Month>4</Month><Day>1</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">24211197</ArticleId><ArticleId IdType="pii">S0006-291X(13)01842-1</ArticleId><ArticleId IdType="doi">10.1016/j.bbrc.2013.10.149</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=IndiumV2/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000797 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 000797 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= *** parameter Area/wikiCode missing ***
|area= IndiumV2
|flux= Main
|étape= Exploration
|type= RBID
|clé= pubmed:24211197
|texte= Detecting ligand interactions with G protein-coupled receptors in real-time on living cells.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:24211197" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \
| NlmPubMed2Wicri -a IndiumV2
| This area was generated with Dilib version V0.5.76. |  |