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Structure and Function of the Bacterial Heterodimeric ABC Transporter CydDC: Stimulation of the ATPase Activity by Thiol and Heme Compounds

Yamashita, Masao, Shepherd, Mark, Booth, Wesley I, Xie, Hao, Postis, Vincent, Nyathi, Yvonne, Tzokov, Svetomir, Poole, Robert K, Baldwin, Stephen A, Bullough, Per A and others. (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 . pp. 23177-23188. ISSN 1083-351X. (doi:10.1074/jbc.M114.590414) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)

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Official URL
http://dx.doi.org/10.1074/jbc.M114.590414

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.

Item Type: Article
DOI/Identification number: 10.1074/jbc.M114.590414
Uncontrolled keywords: ABC Transporter; Bacterial Metabolism; Membrane Protein; Membrane Transporter Reconstitution; Microbiology; Protein Structure; Structural Biology; Transporter
Subjects: Q Science
Q Science > Q Science (General)
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Mark Shepherd
Date Deposited: 07 Jul 2014 09:18 UTC
Last Modified: 23 Jan 2020 04:08 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/41697 (The current URI for this page, for reference purposes)
Shepherd, Mark: https://orcid.org/0000-0002-7472-2300
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