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dc.contributor.authorCranford-Smith, Tamar
dc.contributor.authorJamshad, Mohammed
dc.contributor.authorJeeves, Mark
dc.contributor.authorChandler, Rachael A.
dc.contributor.authorYule, Jack
dc.contributor.authorRobinson, Ashley
dc.contributor.authorAlam, Farhana
dc.contributor.authorDunne, Karl A.
dc.contributor.authorAponte Angarita, Edwin H.
dc.contributor.authorAlanazi, Mashael
dc.contributor.authorCarter, Cailean
dc.contributor.authorHenderson, Ian R.
dc.contributor.authorLovett, Janet E.
dc.contributor.authorWinn, Peter
dc.contributor.authorKnowles, Timothy
dc.contributor.authorHuber, Damon
dc.identifier.citationCranford-Smith , T , Jamshad , M , Jeeves , M , Chandler , R A , Yule , J , Robinson , A , Alam , F , Dunne , K A , Aponte Angarita , E H , Alanazi , M , Carter , C , Henderson , I R , Lovett , J E , Winn , P , Knowles , T & Huber , D 2020 , ' Iron is a ligand of SecA-like metal-binding domains in vivo ' , Journal of Biological Chemistry , vol. Papers in Press .
dc.identifier.otherPURE: 267322114
dc.identifier.otherPURE UUID: a30f7264-b14b-4209-aee5-1761a5cae65c
dc.identifier.otherBibtex: Cranford-Smith02042020
dc.identifier.otherORCID: /0000-0002-3561-450X/work/71955462
dc.identifier.otherWOS: 000537733500028
dc.identifier.otherScopus: 85085258932
dc.descriptionFunding: JEL thanks the Royal Society for a University Research Fellowship and the Wellcome Trust for the Q-band EPR spectrometer (099149/Z/12/Z).en
dc.description.abstractThe ATPase SecA is an essential component of the bacterial Sec machinery, which transports proteins across the cytoplasmic membrane. Most SecA proteins contain a long C-terminal tail (CTT). In Escherichia coli, the CTT contains a structurally flexible linker domain and a small metal-binding domain (MBD). The MBD coordinates zinc via a conserved cysteine-containing motif and binds to SecB and ribosomes. In this study, we screened a high-density transposon library for mutants that affect the susceptibility of E. coli to sodium azide, which inhibits SecA-mediated translocation. Results from sequencing this library suggested that mutations removing the CTT make E. coli less susceptible to sodium azide at subinhibitory concentrations. Copurification experiments suggested that the MBD binds to iron and that azide disrupts iron binding. Azide also disrupted binding of SecA to membranes. Two other E. coli proteins that contain SecA-like MBDs, YecA and YchJ, also copurified with iron, and NMR spectroscopy experiments indicated that YecA binds iron via its MBD. Competition experiments and equilibrium binding measurements indicated that the SecA MBD binds preferentially to iron and that a conserved serine is required for this specificity. Finally, structural modelling suggested a plausible model for the octahedral coordination of iron. Taken together, our results suggest that SecA-like MBDs likely bind to iron in vivo.
dc.relation.ispartofJournal of Biological Chemistryen
dc.rightsCopyright © 2020 the Author(s). Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc. This work has been made available online in accordance with publisher policies or with permission. Permission for further reuse of this content should be sought from the publisher or the rights holder. This is the author created accepted manuscript following peer review and may differ slightly from the final published version. The final published version of this work is available at
dc.subjectSec pathwayen
dc.subjectProtein secretionen
dc.subjectProtein structureen
dc.subjectProtein translocationen
dc.subjectMetal ion-protein interactionen
dc.subjectQC Physicsen
dc.subjectQD Chemistryen
dc.subjectQH301 Biologyen
dc.titleIron is a ligand of SecA-like metal-binding domains in vivoen
dc.typeJournal articleen
dc.contributor.sponsorThe Royal Societyen
dc.contributor.sponsorThe Wellcome Trusten
dc.contributor.institutionUniversity of St Andrews. School of Physics and Astronomyen
dc.contributor.institutionUniversity of St Andrews. Biomedical Sciences Research Complexen
dc.description.statusPeer revieweden

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