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Multiplexed analysi...
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Lorenzen, EmilyRockefeller Univ, Lab Chem Biol & Signal Transduct, 1230 York Ave, New York, NY 10065 USA.
(author)
Multiplexed analysis of the secretin-like GPCR-RAMP interactome
- Article/chapterEnglish2019
Publisher, publication year, extent ...
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AMER ASSOC ADVANCEMENT SCIENCE,2019
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LIBRIS-ID:oai:DiVA.org:kth-263373
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https://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-263373URI
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https://doi.org/10.1126/sciadv.aaw2778DOI
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http://kipublications.ki.se/Default.aspx?queryparsed=id:142118179URI
Supplementary language notes
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Language:English
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Summary in:English
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Subject category:ref swepub-contenttype
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Subject category:art swepub-publicationtype
Notes
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QC 20191115
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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.
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Dodig-Crnkovic, TeaKTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab(Swepub:kth)u1wrth1x
(author)
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Kotliar, Ilana B.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, ElisaKTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab(Swepub:kth)u1pmjyje
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Ceraudo, EmilieRockefeller Univ, Lab Chem Biol & Signal Transduct, 1230 York Ave, New York, NY 10065 USA.
(author)
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Vaughan, Roger D.Rockefeller Univ, Ctr Clin & Translat Sci, 1230 York Ave, New York, NY 10065 USA.
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Uhlén, MathiasKTH,Science for Life Laboratory, SciLifeLab,Systembiologi(Swepub:kth)u1dulvmw
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Huber, ThomasRockefeller Univ, Lab Chem Biol & Signal Transduct, 1230 York Ave, New York, NY 10065 USA.
(author)
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Schwenk, Jochen M.KTH,Science for Life Laboratory, SciLifeLab,Affinitets-proteomik(Swepub:kth)u1h7wtme
(author)
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Sakmar, Thomas P.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)
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In:Science Advances: AMER ASSOC ADVANCEMENT SCIENCE5:92375-2548
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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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Uhlén, Mathias
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Huber, Thomas
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Schwenk, Jochen ...
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Royal Institute of Technology
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Karolinska Institutet