Expression of the <i>Escherichia coli cyo</i> operon in <i>Paracoccus denitrificans</i> results in a fully active quinol oxidase of unexpected heme composition

Schröter, Thomas; Winterstein, Christine; Ludwig, Bernd; Richter, Oliver‐Matthias H.

doi:10.1016/s0014-5793(98)00839-4

Cited by 14 publications

(9 citation statements)

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“…For example, three operons (cyoA, B, C; proV, W, X; and fixA, B, C) are uniquely present in the V. vulnificus small chromosome, and each is tandemly arranged as a single-copy cluster. Genes of the cyo, pro, and fix operons encode different components of cytochrome c oxidase (Schroter et al 1998), the ABCtype proline/glycine betaine transport system (Gowrishankar 1989), and electron transfer flavoprotein, respectively (Weidenhaupt et al 1996). Interestingly, sequences of these operons are related to homologous sequences in Pseudomonas aeruginosa, suggesting that these V. vulnificus-specific sequences might have originated from a distantly related bacterium.…”

Section: Discussion the Evolution Of Vibrio Genomesmentioning

confidence: 99%

Comparative Genome Analysis of Vibrio vulnificus, a Marine Pathogen

et al. 2003

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The halophile Vibrio vulnificus is an etiologic agent of human mortality from seafood-borne infections. We applied whole-genome sequencing and comparative analysis to investigate the evolution of this pathogen. The genome of biotype 1 strain, V. vulnificus YJ016, was sequenced and includes two chromosomes of estimated 3377 kbp and 1857 kbp in size, and a plasmid of 48,508 bp. A super-integron (SI) was identified, and the SI region spans 139 kbp and contains 188 gene cassettes. In contrast to non-SI sequences, the captured gene cassettes are unique for any given Vibrio species and are highly variable among V. vulnificus strains. Multiple rearrangements were found when comparing the 5.3-Mbp V. vulnificus YJ016 genome and the 4.0-Mbp V. cholerae El Tor N16961 genome. The organization of gene clusters of capsular polysaccharide, iron metabolism, and RTX toxin showed distinct genetic features of V. vulnificus and V. cholerae. The content of the V. vulnificus genome contained gene duplications and evidence of horizontal transfer, allowing for genetic diversity and function in the marine environment. The genomic information obtained in this study can be applied to monitoring vibrio infections and identifying virulence genes in V. vulnificus.[Supplemental material is available online at www.genome.org and at http://genome.nhri.org.tw/vv/. The nucleotide sequence data from this study have been submitted to DDBJ/EMBL/GenBank under accession nos. BA000037, BA000038, and AP005352.]Vibrio vulnificus is an etiologic agent for severe human infection acquired through wounds or contaminated seafood. This organism is divided into three biotypes according to their different biochemical and biological properties (Linkous and Oliver 1999). Among them the biotype 1 strains are most frequently isolated from the clinical specimens. Opportunistic infection in susceptible individuals typically causes mortality within 24 to 48 h of the exposure. The bacterium is halophilic, and it is abundantly present in estuarine ecosystems throughout the world. Isolated incidents of V. vulnificus infection have been reported in the U.S.A., Europe, Korea, Taiwan (Park et al. 1991;Chuang et al. 1992;Dalsgaard et al. 1996;Hlady and Klontz 1996), and many other countries. According to CDC statistics, V. vulnificus is a major bacterial cause of mortality associated with food-borne diseases, and it results in the highest death rate of any causative agent (Todd 1989).V. vulnificus belongs to the ␥-group of Proteobacteria, and it shares morphological and biochemical characteristics with other human vibrio pathogens, including Vibrio cholerae and Vibrio parahaemolyticus. Bacteria of the Vibrionaceae family, which show a comma-shape microscopic appearance and a polar flagellum appendage, are mostly aquatic inhabitants that require NaCl for optimal growth. On the basis of clinical and epidemiology studies, diseases associated with V. vulnificus infection have been found to present in two patterns (Blake et al. 1979). In one, primary septicemia occurred in individ...

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Section: Discussion the Evolution Of Vibrio Genomesmentioning

confidence: 99%

Comparative Genome Analysis of Vibrio vulnificus, a Marine Pathogen

et al. 2003

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“…The latter value was based on the assumption that the peak-to-trough absorbance difference is similar for hemes B and O (32). HPLC 1 Heme Analysis-Hemes were extracted from the isolated enzyme complexes essentially as described (33,34). After precipitation with 10 volumes of 90% acetone, 10% water, 10 mM NH 3 , the hemes were extracted from the pellet first into acetone/HCl/water (90:2:8, v/v) and then into diethyl ether.…”

Section: Methodsmentioning

confidence: 99%

Mutation of Arg-54 Strongly Influences Heme Composition and Rate and Directionality of Electron Transfer in Paracoccus denitrificans Cytochrome c Oxidase

Kannt¹,

Pfitzner²,

Ruitenberg³

et al. 1999

Journal of Biological Chemistry

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The effect of a single site mutation of Arg-54 to methionine in Paracoccus denitrificans cytochrome c oxidase was studied using a combination of optical spectroscopy, electrochemical and rapid kinetics techniques, and time-resolved measurements of electrical membrane potential. The mutation resulted in a blue-shift of the heme a ␣-band by 15 nm and partial occupation of the low-spin heme site by heme O. Additionally, there was a marked decrease in the midpoint potential of the low-spin heme, resulting in slow reduction of this heme species. A stopped-flow investigation of the reaction with ferrocytochrome c yielded a kinetic difference spectrum resembling that of heme a 3 . This observation, and the absence of transient absorbance changes at the corresponding wavelength of the low-spin heme, suggests that, in the mutant enzyme, electron transfer from Cu A to the binuclear center may not occur via heme a but that instead direct electron transfer to the high-spin heme is the dominating process. This was supported by charge translocation measurements where ⌬ generation was completely inhibited in the presence of KCN. Our results thus provide an example for how the interplay between protein and cofactors can modulate the functional properties of the enzyme complex.

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“…16 Wild-type bo 3 enzyme with bound UQ 8 was purified as described previously. 29 Native UQ 8 was removed by purification of QOX using N,N-dimethyldodecylamine N-oxide (LDAO) as the detergent, followed by detergent exchange with n-dodecyl-b-D-maltoside (b-DM). Reconstitution with exogenous 2,3-dimethoxy-5,6-dimethyl-1,4-benzoquinone (DDQ, see Fig.…”

Section: Sample Preparationmentioning

confidence: 99%

Elucidating mechanisms in haemcopperoxidases: The high-affinity Q_Hbinding site in quinol oxidase as studied by DONUT-HYSCOREspectroscopy and density functional theory

et al. 2011

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Expression of the Escherichia coli cyo operon in Paracoccus denitrificans results in a fully active quinol oxidase of unexpected heme composition

Cited by 14 publications

References 30 publications

Comparative Genome Analysis of Vibrio vulnificus, a Marine Pathogen

Comparative Genome Analysis of Vibrio vulnificus, a Marine Pathogen

Mutation of Arg-54 Strongly Influences Heme Composition and Rate and Directionality of Electron Transfer in Paracoccus denitrificans Cytochrome c Oxidase

Elucidating mechanisms in haemcopperoxidases: The high-affinity Q_Hbinding site in quinol oxidase as studied by DONUT-HYSCOREspectroscopy and density functional theory

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Expression of the Escherichia coli cyo operon in Paracoccus denitrificans results in a fully active quinol oxidase of unexpected heme composition

Cited by 14 publications

References 30 publications

Comparative Genome Analysis of Vibrio vulnificus, a Marine Pathogen

Comparative Genome Analysis of Vibrio vulnificus, a Marine Pathogen

Mutation of Arg-54 Strongly Influences Heme Composition and Rate and Directionality of Electron Transfer in Paracoccus denitrificans Cytochrome c Oxidase

Elucidating mechanisms in haemcopperoxidases: The high-affinity QHbinding site in quinol oxidase as studied by DONUT-HYSCOREspectroscopy and density functional theory

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Elucidating mechanisms in haemcopperoxidases: The high-affinity Q_Hbinding site in quinol oxidase as studied by DONUT-HYSCOREspectroscopy and density functional theory