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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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- 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.
- In: Science Advances. - : AMER ASSOC ADVANCEMENT SCIENCE. - 2375-2548. ; 5:9
- Journal article (peer-reviewed)
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)
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