Heterologous production in Wolinella succinogenes and characterization of the quinol : fumarate reductase enzymes from Helicobacter pylori and Campylobacter jejuni

Mileni, Mauro, Macmillan, Fraser, Tziatzios, Christos, Zwicker, Klaus, Haas, Alexander H., Mantele, Werner, Simon, Jorg and Lancaster, C. Roy D. (2006) Heterologous production in Wolinella succinogenes and characterization of the quinol : fumarate reductase enzymes from Helicobacter pylori and Campylobacter jejuni. Biochemical Journal, 395 (1). pp. 191-201. ISSN 0264-6021

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Abstract

The epsilon-proteobacteria Helicobacter pylori and Campylobacter jejuni are both human pathogens. They colonize mucosal surfaces causing severe diseases. The membrane protein complex QFR (quinol:fumarate reductase) from H. pylori has previously been established as a potential drug target, and the same is likely for the QFR from C. jejuni. In the present paper, we describe the cloning of the QFR operons from the two pathogenic bacteria H. pylori and C. jejuni and their expression in Wolinella succinogenes, a non-pathogenic epsilon-proteobacterium. To our knowledge, this is the first documentation of heterologous membrane protein production in W succinogenes. We demonstrate that the replacement of the homologous enzyme from W succinogenes with the heterologous enzymes yields Mutants where fumarate respiration is fully functional. We have isolated and characterized the heterologous QFR enzymes. The high quality of the enzyme preparation enabled us to determine unequivocally by analytical ultracentrifugation the homodimeric state of the three detergent-solubilized heterotrimeric QFR enzymes, to accurately determine the different oxidation-reduction ('redox') midpoint potentials of the six prosthetic groups, the Michaelis constants for the quinol substrate, maximal enzymatic activities and the characterization of three different anti-helminths previously suggested to be inhibitors of the QFR enzymes from H. pylori and C. jejuni. This characterization allows, for the first time, a detailed comparison of the QFR enzymes from C. jejuni and H. pylori with that of W. succinogenes.

Item Type: Article
Faculty \ School: Faculty of Science > School of Chemistry
Depositing User: Rachel Smith
Date Deposited: 18 Jul 2011 15:49
Last Modified: 21 Apr 2020 19:49
URI: https://ueaeprints.uea.ac.uk/id/eprint/33897
DOI: 10.1042/bj20051675

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