Yamashita, M, Shepherd, M, Booth, WI et al. (7 more authors) (2014) Structure and Function of the Bacterial Heterodimeric ABC Transporter CydDC: Stimulation of the ATPase Activity by Thiol and Heme Compounds. The Journal of Biological Chemistry, 289 (33). pp. 23177-23188. ISSN 0021-9258
Abstract
In Escherichia coli, the biogenesis of both cytochrome bd-type quinol oxidases and periplasmic cytochromes requires the ABC- type cysteine/glutathione (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 analysed by electron cryomicrosopy 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 ABC 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 (ca. 100 nmol Pi/min/mg) that was stimulated approximately three-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.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | (c) 2014 by The American Society for Biochemistry and Molecular Biology, Inc. This is an open access article under the terms of the Creative Commons Attribution License CC-BY-3.0. (https://creativecommons.org/licenses/by/3.0/) |
Keywords: | ABC Transporter; Bacterial Metabolism; Membrane Protein; Membrane Transporter Reconstitution; Microbiology; Protein Structure; Structural Biology; Transporter |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biomedical Sciences (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 19 Mar 2018 16:17 |
Last Modified: | 25 Jun 2023 21:12 |
Status: | Published |
Publisher: | American Society for Biochemistry and Molecular Biology Inc. |
Identification Number: | 10.1074/jbc.M114.590414 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:126457 |