Show simple item record

dc.contributor.authorMajd, Homa
dc.contributor.authorKing, Martin S
dc.contributor.authorPalmer, Shane M
dc.contributor.authorSmith, Anthony C
dc.contributor.authorElbourne, Liam Dh
dc.contributor.authorPaulsen, Ian T
dc.contributor.authorSharples, David
dc.contributor.authorHenderson, Peter Jf
dc.contributor.authorKunji, Edmund Rs
dc.date.accessioned2018-11-23T00:31:34Z
dc.date.available2018-11-23T00:31:34Z
dc.date.issued2018-10-15
dc.identifier.issn2050-084X
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/285785
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.format.mediumElectronic
dc.languageeng
dc.publishereLife Sciences Publications, Ltd
dc.rightsAttribution 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectMitochondria
dc.subjectAnimals
dc.subjectHumans
dc.subjectTetrahymena
dc.subjectIons
dc.subjectMembrane Transport Proteins
dc.subjectAdenosine Diphosphate
dc.subjectAdenosine Triphosphate
dc.subjectLigands
dc.subjectBiological Assay
dc.subjectReproducibility of Results
dc.subjectTemperature
dc.subjectSubstrate Specificity
dc.subjectProtein Stability
dc.titleScreening of candidate substrates and coupling ions of transporters by thermostability shift assays.
dc.typeArticle
prism.publicationDate2018
prism.publicationNameElife
prism.volume7
dc.identifier.doi10.17863/CAM.33129
dcterms.dateAccepted2018-10-11
rioxxterms.versionofrecord10.7554/eLife.38821
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2018-10-15
dc.contributor.orcidMajd, Homa [0000-0002-2048-1839]
dc.contributor.orcidKing, Martin S [0000-0001-6030-5154]
dc.contributor.orcidHenderson, Peter Jf [0000-0002-9187-0938]
dc.contributor.orcidKunji, Edmund Rs [0000-0002-0610-4500]
dc.identifier.eissn2050-084X
rioxxterms.typeJournal Article/Review
pubs.funder-project-idMedical Research Council (MC_U105663139)
pubs.funder-project-idMedical Research Council (MC_UU_00015/1)
cam.issuedOnline2018-10-15
cam.orpheus.successThu Jan 30 10:53:57 GMT 2020 - The item has an open VoR version.
rioxxterms.freetoread.startdate2100-01-01


Files in this item

Thumbnail
Thumbnail

This item appears in the following Collection(s)

Show simple item record

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