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Multiplexed analysi...
Multiplexed analysis of the secretin-like GPCR-RAMP interactome
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- Lorenzen, Emily (author)
- Rockefeller Univ, Lab Chem Biol & Signal Transduct, 1230 York Ave, New York, NY 10065 USA.
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- Dodig-Crnkovic, Tea (author)
- KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab
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- Kotliar, Ilana B. (author)
- Rockefeller Univ, Lab Chem Biol & Signal Transduct, 1230 York Ave, New York, NY 10065 USA.;Tri Inst PhD Program Chem Biol, New York, NY 10065 USA.
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- Pin, Elisa (author)
- KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab
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- Ceraudo, Emilie (author)
- Rockefeller Univ, Lab Chem Biol & Signal Transduct, 1230 York Ave, New York, NY 10065 USA.
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- Vaughan, Roger D. (author)
- Rockefeller Univ, Ctr Clin & Translat Sci, 1230 York Ave, New York, NY 10065 USA.
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- Uhlén, Mathias (author)
- KTH,Science for Life Laboratory, SciLifeLab,Systembiologi
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- Huber, Thomas (author)
- Rockefeller Univ, Lab Chem Biol & Signal Transduct, 1230 York Ave, New York, NY 10065 USA.
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- Schwenk, Jochen M. (author)
- KTH,Science for Life Laboratory, SciLifeLab,Affinitets-proteomik
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- Sakmar, Thomas P. (author)
- Karolinska Institutet
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Rockefeller Univ, Lab Chem Biol & Signal Transduct, 1230 York Ave, New York, NY 10065 USA Affinitets-proteomik (creator_code:org_t)
- AMER ASSOC ADVANCEMENT SCIENCE, 2019
- 2019
- English.
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In: Science Advances. - : AMER ASSOC ADVANCEMENT SCIENCE. - 2375-2548. ; 5:9
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https://urn.kb.se/re...
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Abstract
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- 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.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Lorenzen, Emily
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Dodig-Crnkovic, ...
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Kotliar, Ilana B ...
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Pin, Elisa
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Ceraudo, Emilie
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Vaughan, Roger D ...
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show more...
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Uhlén, Mathias
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Huber, Thomas
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Schwenk, Jochen ...
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Sakmar, Thomas P ...
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- About the subject
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Basic Medicine
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and Cell and Molecul ...
- Articles in the publication
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Science Advances
- By the university
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Royal Institute of Technology
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Karolinska Institutet