In:
Journal of Bacteriology, American Society for Microbiology, Vol. 189, No. 13 ( 2007-07), p. 4764-4773
Kurzfassung:
Pyruvate-dependent reduction of NADP has been demonstrated in cell extracts of the human gastric pathogen Helicobacter pylori. However, NADP is not a substrate of purified pyruvate:ferredoxin oxidoreductase (PFOR), suggesting that other redox active enzymes mediate this reaction. Here we show that fqrB (HP1164), which is essential and highly conserved among the epsilonproteobacteria, exhibits NADPH oxidoreductase activity. FqrB was purified by nickel interaction chromatography following overexpression in Escherichia coli . The protein contained flavin adenine dinucleotide and exhibited NADPH quinone reductase activity with menadione or benzoquinone and weak activity with cytochrome c , molecular oxygen, and 5,5′-dithio-bis-2-nitrobenzoic acid (DTNB). FqrB exhibited a ping-pong catalytic mechanism, a k cat of 122 s −1 , and an apparent K m of 14 μM for menadione and 26 μM for NADPH. FqrB also reduced flavodoxin (FldA), the electron carrier of PFOR. In coupled enzyme assays with purified PFOR and FldA, FqrB reduced NADP in a pyruvate- and reduced coenzyme A (CoA)-dependent manner. Moreover, in the presence of NADPH, CO 2 , and acetyl-CoA, the PFOR:FldA:FqrB complex generated pyruvate via CO 2 fixation. PFOR was the rate-limiting enzyme in the complex, and nitazoxanide, a specific inhibitor of PFOR of H. pylori and Campylobacter jejuni , also inhibited NADP reduction in cell-free lysates. These capnophilic (CO 2 -requiring) organisms contain gaps in pathways of central metabolism that would benefit substantially from pyruvate formation via CO 2 fixation. Thus, FqrB provides a novel function in pyruvate metabolism and, together with production of superoxide anions via quinone reduction under high oxygen tensions, contributes to the unique microaerobic lifestyle that defines the epsilonproteobacterial group.
Materialart:
Online-Ressource
ISSN:
0021-9193
,
1098-5530
Sprache:
Englisch
Verlag:
American Society for Microbiology
Publikationsdatum:
2007
ZDB Id:
1481988-0
SSG:
12
