Structure and function of the bacterial heterodimeric ABC transporter CydDC: stimulation of ATPase activity by thiol and heme compounds.
dc.contributor.author | Yamashita, M. | |
dc.contributor.author | Shepherd, M. | |
dc.contributor.author | Booth, W.I. | |
dc.contributor.author | Xie, H. | |
dc.contributor.author | Postis, V. | |
dc.contributor.author | Nyathi, Yvonne | |
dc.contributor.author | Tzokov, S.B. | |
dc.contributor.author | Poole, R.K. | |
dc.contributor.author | Baldwin, S.A. | |
dc.contributor.author | Bullough, P.A. | |
dc.date.accessioned | 2020-06-10T07:47:08Z | |
dc.date.accessioned | 2020-07-08T13:39:25Z | |
dc.date.available | 2020-06-10T07:47:08Z | |
dc.date.available | 2020-07-08T13:39:25Z | |
dc.date.issued | 2014-08 | |
dc.identifier.citation | Yamashita 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.uri | http://hdl.handle.net/10454/17911 | |
dc.description | Yes | en_US |
dc.description.abstract | In 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.sponsorship | This work was supported by Biotechnology and Biological Sciences Research Council grant BBS/B/14418 (Membrane Protein Structure Initiative). | en_US |
dc.language.iso | en | en_US |
dc.relation.isreferencedby | https://doi.org/10.1074/jbc.M114.590414 | en_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.subject | ABC Transporter | en_US |
dc.subject | Bacterial metabolism | en_US |
dc.subject | Membrane protein | en_US |
dc.subject | Membrane transporter reconstitution | en_US |
dc.subject | Microbiology | en_US |
dc.subject | Protein structure | en_US |
dc.subject | Structural biology | en_US |
dc.subject | Transporter | en_US |
dc.title | Structure and function of the bacterial heterodimeric ABC transporter CydDC: stimulation of ATPase activity by thiol and heme compounds. | en_US |
dc.status.refereed | Yes | en_US |
dc.date.Accepted | 2014-06 | |
dc.date.application | 2014-06-23 | |
dc.type | Article | en_US |
dc.type.version | Published version | en_US |
dc.date.updated | 2020-06-10T06:47:08Z | |
refterms.dateFOA | 2020-07-08T13:40:17Z |