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dc.contributor.authorMajd, Homaen
dc.contributor.authorKing, Martinen
dc.contributor.authorPalmer, Shaneen
dc.contributor.authorSmith, Anthonyen
dc.contributor.authorElbourne, Liamen
dc.contributor.authorPaulsen, Ianen
dc.contributor.authorSharples, Daviden
dc.contributor.authorHenderson, Peteren
dc.contributor.authorKunji, Edmund RSen
dc.date.accessioned2019-01-08T00:31:35Z
dc.date.available2019-01-08T00:31:35Z
dc.date.issued2018-10-12en
dc.identifier.issn2050-084X
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/287627
dc.description.abstractSubstrates of most transport proteins have not been identified, limiting our understanding of their role in physiology and disease. Traditional identification methods use transport assays with radioactive compounds, but they are technically challenging and many compounds are unavailable in radioactive form or are prohibitively expensive, precluding large-scale trials. Here, we present a high-throughput screening method that can identify candidate substrates from libraries of unlabeled compounds. The assay is based on the principle that transport proteins recognize substrates through specific interactions, which lead to enhanced stabilization of the transporter population in thermostability shift assays. Representatives of three different transporter (super)families were tested, which differ in structure as well as transport and ion coupling mechanisms. In each case, the substrates were identified correctly from a large set of chemically related compounds, including stereo-isoforms. In some cases, stabilization by substrate binding was enhanced further by ions, providing testable hypotheses on energy coupling mechanisms.
dc.publishereLife Sciences Publications Ltd
dc.rightsAttribution 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.titleScreening of candidate substrates and coupling ions of transporters by thermostability shift assaysen
dc.typeArticle
prism.publicationDate2018en
prism.publicationNameeLifeen
dc.identifier.doi10.17863/CAM.34939
dcterms.dateAccepted2018-10-12en
rioxxterms.versionofrecord10.7554/eLife.38821en
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2018-10-12en
dc.contributor.orcidKing, Martin [0000-0001-6030-5154]
dc.contributor.orcidSmith, Anthony [0000-0003-0141-0434]
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idMRC (MC_U105663139)
cam.issuedOnline2018-10-15en


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International