Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Multiplexed analysis of the secretin-like GPCR-RAMP interactome
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Affinity Proteomics. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0002-2875-896x
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Affinity Proteomics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
Show others and affiliations
2019 (English)In: Science Advances, E-ISSN 2375-2548, Vol. 5, no 9, article id eaaw2778Article in journal (Refereed) Published
Abstract [en]

Receptor activity–modifying proteins (RAMPs) have been shown to modulate the functions of several G protein–coupled receptors (GPCRs), but potential direct interactions among the three known RAMPs and hundreds of GPCRs have never been investigated. Focusing mainly on the secretin-like family of GPCRs, we engineered epitope-tagged GPCRs and RAMPs, and developed a multiplexed suspension bead array (SBA) immunoassay to detect GPCR-RAMP complexes from detergent-solubilized lysates. Using 64 antibodies raised against the native proteins and 4 antibodies targeting the epitope tags, we mapped the interactions among 23 GPCRs and 3 RAMPs. We validated nearly all previously reported secretin-like GPCR-RAMP interactions, and also found previously unidentified RAMP interactions with additional secretin-like GPCRs, chemokine receptors, and orphan receptors. The results provide a complete interactome of secretin-like GPCRs with RAMPs. The SBA strategy will be useful to search for additional GPCR-RAMP complexes and other interacting membrane protein pairs in cell lines and tissues. Copyright

Place, publisher, year, edition, pages
American Association for the Advancement of Science , 2019. Vol. 5, no 9, article id eaaw2778
Keywords [en]
Antibodies, Cell culture, Chemokine receptors, Coupled receptors, Direct interactions, Membrane proteins, Multiplexed analysis, Native proteins, Receptor activity, Suspension bead arrays, Epitopes
National Category
Industrial Biotechnology
Research subject
Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-263565DOI: 10.1126/sciadv.aaw2778ISI: 000491128800060PubMedID: 31555726Scopus ID: 2-s2.0-85072340840OAI: oai:DiVA.org:kth-263565DiVA, id: diva2:1371558
Note

QC20191120

Available from: 2019-11-20 Created: 2019-11-20 Last updated: 2020-01-10Bibliographically approved

Open Access in DiVA

No full text in DiVA

Other links

Publisher's full textPubMedScopus

Authority records BETA

Dodig-Crnkovic, TeaUhlén, MathiasSchwenk, Jochen M.

Search in DiVA

By author/editor
Dodig-Crnkovic, TeaPin, ElisaUhlén, MathiasSchwenk, Jochen M.
By organisation
Affinity ProteomicsScience for Life Laboratory, SciLifeLabSystems Biology
In the same journal
Science Advances
Industrial Biotechnology

Search outside of DiVA

GoogleGoogle Scholar

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 35 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf