Production, characterization and determination of the real catalytic properties of the putative "succinate dehydrogenase" from Wolinella succinogenes
- Both the genomes of the epsilonproteobacteria Wolinella succinogenes and Campylobacter jejuni contain operons (sdhABE) that encode for so far uncharacterized enzyme complexes annotated as ‘non-classical’ succinate:quinone reductases (SQRs). However, the role of such an enzyme ostensibly involved in aerobic respiration in an anaerobic organism such as W. succinogenes has hitherto been unknown. We have established the first genetic system for the manipulation and production of a member of the non-classical succinate:quinone oxidoreductase family. Biochemical characterization of the W. succinogenes enzyme reveals that the putative SQR is in fact a novel methylmenaquinol:fumarate reductase (MFR) with no detectable succinate oxidation activity, clearly indicative of its involvement in anaerobic metabolism. We demonstrate that the hydrophilic subunits of the MFR complex are, in contrast to all other previously characterized members of the superfamily, exported into the periplasm via the twin-arginine translocation (tat)-pathway. Furthermore we show that a single amino acid exchange (Ala86→His) in the flavoprotein of that enzyme complex is the only additional requirement for the covalent binding of the otherwise non-covalently bound FAD. Our results provide an explanation for the previously published puzzling observation that the C. jejuni sdhABE operon is upregulated in an oxygen-limited environment as compared with microaerophilic laboratory conditions.
Author: | Hanno Dominik Juhnke, Heiko Hiltscher, Hamid Reza Nasiri, Harald SchwalbeORCiDGND, C. Roy D. LancasterORCiDGND |
---|---|
URN: | urn:nbn:de:hebis:30:3-272782 |
DOI: | https://doi.org/10.1111/j.1365-2958.2008.06581.x |
ISSN: | 1088–1101 |
ISSN: | 1365-2958 |
Parent Title (English): | Molecular microbiology |
Publisher: | Wiley-Blackwell |
Place of publication: | Oxford [u.a.] |
Document Type: | Article |
Language: | English |
Date of Publication (online): | 2012/12/18 |
Date of first Publication: | 2009/01/16 |
Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
Release Date: | 2012/12/18 |
Volume: | 71 |
Issue: | 5 |
Page Number: | 14 |
First Page: | 1088 |
Last Page: | 1101 |
Note: | Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation. |
HeBIS-PPN: | 316073741 |
Institutes: | Biochemie, Chemie und Pharmazie / Biochemie und Chemie |
Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie |
Sammlungen: | Universitätspublikationen |
Sammlung Biologie / Sondersammelgebiets-Volltexte | |
Licence (German): | Creative Commons - Namensnennung-Nicht kommerziell 3.0 |