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dc.contributor.authorYamashita, M.
dc.contributor.authorShepherd, M.
dc.contributor.authorBooth, W.I.
dc.contributor.authorXie, H.
dc.contributor.authorPostis, V.
dc.contributor.authorNyathi, Yvonne
dc.contributor.authorTzokov, S.B.
dc.contributor.authorPoole, R.K.
dc.contributor.authorBaldwin, S.A.
dc.contributor.authorBullough, P.A.
dc.date.accessioned2020-06-10T07:47:08Z
dc.date.accessioned2020-07-08T13:39:25Z
dc.date.available2020-06-10T07:47:08Z
dc.date.available2020-07-08T13:39:25Z
dc.date.issued2014-08
dc.identifier.citationYamashita M, Shepherd M, Booth WI et al (2014) Structure and function of the bacterial heterodimeric ABC transporter CydDC: stimulation of ATPase activity by thiol and heme compounds. Journal of Biological Chemistry. 289(33). 23177-23188.en_US
dc.identifier.urihttp://hdl.handle.net/10454/17911
dc.descriptionYesen_US
dc.description.abstractIn Escherichia coli, the biogenesis of both cytochrome bd-type quinol oxidases and periplasmic cytochromes requires the ATP-binding cassette-type cysteine/GSH transporter, CydDC. Recombinant CydDC was purified as a heterodimer and found to be an active ATPase both in soluble form with detergent and when reconstituted into a lipid environment. Two-dimensional crystals of CydDC were analyzed by electron cryomicroscopy, and the protein was shown to be made up of two non-identical domains corresponding to the putative CydD and CydC subunits, with dimensions characteristic of other ATP-binding cassette transporters. CydDC binds heme b. Detergent-solubilized CydDC appears to adopt at least two structural states, each associated with a characteristic level of bound heme. The purified protein in detergent showed a weak basal ATPase activity (approximately 100 nmol Pi/min/mg) that was stimulated ∼3-fold by various thiol compounds, suggesting that CydDC could act as a thiol transporter. The presence of heme (either intrinsic or added in the form of hemin) led to a further enhancement of thiol-stimulated ATPase activity, although a large excess of heme inhibited activity. Similar responses of the ATPase activity were observed with CydDC reconstituted into E. coli lipids. These results suggest that heme may have a regulatory role in CydDC-mediated transmembrane thiol transport.en_US
dc.description.sponsorshipThis work was supported by Biotechnology and Biological Sciences Research Council grant BBS/B/14418 (Membrane Protein Structure Initiative).en_US
dc.language.isoenen_US
dc.relation.isreferencedbyhttps://doi.org/10.1074/jbc.M114.590414en_US
dc.rights© 2014 by The American Society for Biochemistry and Molecular Biology, Inc. Creative Commons Attribution Unported License (https://creativecommons.org/licenses/by/3.0/)en_US
dc.subjectABC Transporteren_US
dc.subjectBacterial metabolismen_US
dc.subjectMembrane proteinen_US
dc.subjectMembrane transporter reconstitutionen_US
dc.subjectMicrobiologyen_US
dc.subjectProtein structureen_US
dc.subjectStructural biologyen_US
dc.subjectTransporteren_US
dc.titleStructure and function of the bacterial heterodimeric ABC transporter CydDC: stimulation of ATPase activity by thiol and heme compounds.en_US
dc.status.refereedYesen_US
dc.date.Accepted2014-06
dc.date.application2014-06-23
dc.typeArticleen_US
dc.type.versionPublished versionen_US
dc.date.updated2020-06-10T06:47:08Z
refterms.dateFOA2020-07-08T13:40:17Z


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