DOCSLIB.ORG

Aerobic Respiratory Chain of Escherichia Coli Is Not Allowed to Work in Fully Uncoupled Mode

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Aerobic Respiratory Chain of Escherichia Coli Is Not Allowed to Work in Fully Uncoupled Mode Aerobic respiratory chain of Escherichia coli is not allowed to work in fully uncoupled mode Vitaliy B. Borisova,1, Ranjani Muralib,1, Marina L. Verkhovskayac, Dmitry A. Blochc, Huazhi Hanb, Robert B. Gennisb, and Michael I. Verkhovskyc,2 aBelozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119991, Russia; bDepartment of Biochemistry, University of Illinois, Urbana, IL 61801; and cHelsinki Bioenergetics Group, Institute of Biotechnology, University of Helsinki, PB 65 (Viikinkaari 1), FI-00014 Helsinki, Finland Edited* by Pierre A. Joliot, Institut de Biologie Physico-Chimique, Paris, France, and approved August 11, 2011 (received for review May 26, 2011) Escherichia coli is known to couple aerobic respiratory catabolism to ATP synthesis by virtue of the primary generators of the proton — bo motive force NADH dehydrogenase I, cytochrome 3, and cyto- chrome bd-I. An E. coli mutant deficient in NADH dehydrogenase I, bo bd 3 and -I can, nevertheless, grow aerobically on nonfermenta- ble substrates, although its sole terminal oxidase cytochrome bd-II has been reported to be nonelectrogenic. In the current work, the ability of cytochrome bd-II to generate a proton motive force is reexamined. Absorption and fluorescence spectroscopy and oxy- gen pulse methods show that in the steady-state, cytochrome Fig. 1. Scheme of the components of the E. coli aerobic respiratory bd-II does generate a proton motive force with a Hþ∕e− ratio of chain, starting with NADH as the substrate. Enzyme bioenergetic efficiency 0.94 Æ 0.18. This proton motive force is sufficient to drive ATP is indicated as the number of protons released into the periplasm per elec- tron (Hþ∕e− ratio). NDH-I and NDH-II are the coupled and uncoupled NADH: synthesis and transport of nutrients. Microsecond time-resolved, quinone oxidoreductases, respectively, and UQ-8 is ubiquinone-8. NDH-I and single-turnover electrometry shows that the molecular mechanism NDH-II transfer electrons to UQ-8 to yield reduced UQ-8. Three quinol:oxygen BIOCHEMISTRY of generating the proton motive force is identical to that in cyto- oxidoreductases, cytochromes bo3, bd-I, bd-II, oxidize reduced UQ-8 and chrome bd-I. The ability to induce cytochrome bd-II biosynthesis reduce O2 to 2 H2O. allows E. coli to remain energetically competent under a variety of environmental conditions. ginating from opposite sides of the membrane to generate water ðHþ∕e− ¼ 1Þ (5, 8–11). In addition, there are a number of erobic bacteria produce ATP via (i) oxygen-independent, substrate dehydrogenases that are not electrogenic, including Asubstrate-level phosphorylation in glycolysis and the Krebs the nonelectrogenic NADH:quinone oxidoreductase, NDH-II cycle, or (ii) oxidative phosphorylation, in which the electron ðHþ∕e− ¼ 0Þ. flow, through the aerobic respiratory electron transport chain, Under growth conditions with high aeration, cytochrome bo3 generates a proton motive force ðΔp%Þ which, in turn, drives predominates, whereas, under microaerophilic conditions, cyto- the ATP synthase. The proton motive force (Δpout-in, where it chrome bd-I is the predominant respiratory oxygen reductase that is specified that the gradient is measured “outside-minus-inside”) is present. Conditions of carbon and phosphate starvation result is equivalent to the transmembrane electrochemical proton in the induction of a third respiratory oxygen reductase, cyto- gradient and has chemical ðΔpHout-inÞ and electrical ðΔΨout-inÞ chrome bd-II encoded by appBC genes (also called cyxAB or components. At 298 K, in mV units, cbdAB) (12–14). The ability of cytochrome bd-II to generate a proton motive force is addressed in the current work. Δpout-in ¼ ΔΨout-in − 59ΔpHout-in Whereas cytochrome bo3 and cytochrome bd-I have been extensively studied, very little is known about cytochrome bd-II. For bacteria, such as Escherichia coli, grown at neutral pH, the Cytochrome bd-I is a two-subunit enzyme carrying three hemes: major component of the proton motive force is ΔΨout-in, which is b558, b595, and d (15–17). The latter two hemes are suggested positive outside and typically has a value in the range of 100 to to compose a unique di-heme site for the capture of O2 and 180 mV. Bacteria usually possess branched respiratory chains in its reduction to H2O. Heme d plays the major role in O2 binding which the different components are induced to allow the cells to and formation of oxygenated intermediates (9, 18–25). In con- adapt to various environmental conditions (1). One of the para- trast, cytochrome bd-II remains poorly studied, but it has been meters that depends on the composition of the respiratory chain shown that its spectral properties closely resemble those of cyto- is the “bioenergetic efficiency,” which can be defined as number chrome bd-I (26). The amino acid sequences of both subunits of charges driven across the membrane per electron used to of cytochrome bd-I and bd-II are also highly homologous (about reduce oxygen to water, ðq∕e−Þ. In the E. coli respiratory chain, 60% identity). the net charge transfer across the membrane is equal to the num- It has been reported recently that cytochrome bd-II does not ber of protons released into the periplasm ðHþ∕e−Þ, which can be contribute to the generation of the proton motive force ð þ∕ − ¼ 0Þ measured. H e (27). This was deduced from the growth properties The aerobic respiratory chain of E. coli (Fig. 1) contains three i enzymes that are known to generate a proton motive force: ( ) the Author contributions: V.B.B., M.L.V., D.A.B., R.B.G., and M.I.V. designed research; V.B.B., proton-pumping NADH:quinone oxidoreductase NADH dehy- R.M., M.L.V., D.A.B., H.H., and M.I.V. performed research; M.I.V. contributed new þ − drogenase I (NDH-I) ðH ∕e ¼ 2Þ (2); (ii) cytochrome bo3 reagents/analytic tools; V.B.B., R.M., M.L.V., D.A.B., H.H., R.B.G., and M.I.V. analyzed data; ðHþ∕e− ¼ 2Þ (3), the proton-pumping oxygen reductase that is and V.B.B., M.L.V., D.A.B., R.B.G., and M.I.V. wrote the paper. a member of the superfamily of heme-copper oxidoreductases The authors declare no conflict of interest. (4–7); and (iii) cytochrome bd-I, which is unable to pump protons *This Direct Submission article had a prearranged editor. but generates a proton motive force by transmembrane charge 1V.B.B. and R.M. contributed equally to this work. separation resulting from utilizing protons and electrons ori- 2To whom correspondence should be addressed. E-mail: [email protected]. www.pnas.org/cgi/doi/10.1073/pnas.1108217108 PNAS Early Edition ∣ 1of5 Downloaded by guest on September 23, 2021 of the strain that was genetically constructed to lack NDH-I, data show that ΔΨ across the bacterial membrane is generated cytochrome bo3 and cytochrome bd-I. The aerobic respiratory due to either turnover of the F1FO ATPase (trace 1) or by quinol chain from NADH is constrained to NDH-II → bd-II. By exam- oxidase activity (trace 2). Because the only quinol oxidase present ination of the growth of this strain under glucose-limited condi- is cytochrome bd-II, these data imply that catalytic turnover of tions in continuous culture, it was concluded that cytochrome cytochrome bd-II generates a transmembrane potential (ΔΨ) bd-II does not contribute to the proton motive force (i.e., the under steady-state conditions. aerobic respiratory chain is fully uncoupled). Shepherd et al. (28) Fig. 2B shows the responses of the dye acridine orange (AO) have proposed that under such conditions E. coli creates the under the same conditions, reporting the internal acidification proton motive force by means of consumption of intracellular in the inverted vesicles, and, therefore, formation of ΔpH. The γ protons during the synthesis of -aminobutyric acid (GABA), addition of either ATP (trace 1) or Q1 plus DTT (trace 2) results which is coupled to the electrogenic uptake of anionic glutamate in quenching of the AO fluorescence, demonstrating acidification by the glutamate/GABA antiporter. of the vesicle interior. These pH changes are abolished by grami- In this work we directly determined the ability of cytochrome cidin. The data clearly show that catalytic turnover of cytochrome bd -II to generate a proton motive force by examining intact bd-II is accompanied by the release of protons on the periplasmic cells, spheroplasts, and membrane vesicles as well as purified side of the membrane, equivalent to the inside of the inverted bd cytochrome -II. The data definitively show that the catalytic vesicles. bd turnover of cytochrome -II does generate a proton motive It is noted that the observed kinetics of the generation of ΔΨ bd force by a mechanism identical to that of cytochrome -I. A and ΔpH are different (Fig. 2). This is because ΔΨ is produced bd previous proposal (29) that the -type oxygen reductase from almost immediately, whereas ΔpH develops at a much slower rate Azotobacter vinlandii is not coupled has also been demonstrated since the membrane vesicles are loaded with concentrated buffer. not to be correct (30). It is very likely, therefore, that all members bd of the family of -type oxygen reductases generate a proton Hþ∕e− Ratio Measurement. Fig. 3B shows the change in the pH of motive force even as they may have additional roles such as an anaerobic suspension of spheroplasts from E. coli MB37 (lack- scavenging O2 or maintaining redox balance in the cytoplasm. ing NDH-I, cytochrome bo3 and cytochrome bd-I) upon addition The data presented has import in examining the bioenergetics of aliquots of O2.
Load more

Recommended publications
  • Bacterial Oxidases of the Cytochrome Bd Family: Redox Enzymes of Unique Structure, Function, and Utility As Drug Targets
    Published in "Antioxidants & Redox Signaling doi: 10.1089/ars.2020.8039, 2020" which should be cited to refer to this work. Bacterial Oxidases of the Cytochrome bd Family: Redox Enzymes of Unique Structure, Function, and Utility As Drug Targets Vitaliy B. Borisov,1 Sergey A. Siletsky,1 Alessandro Paiardini,2 David Hoogewijs,3 Elena Forte,2 Alessandro Giuffre`,4 and Robert K. Poole5 Abstract Significance: Cytochrome bd is a ubiquinol:oxygen oxidoreductase of many prokaryotic respiratory chains with a unique structure and functional characteristics. Its primary role is to couple the reduction of molecular oxygen, even at submicromolar concentrations, to water with the generation of a proton motive force used for adenosine triphosphate production. Cytochrome bd is found in many bacterial pathogens and, surprisingly, in bacteria for- mally denoted as anaerobes. It endows bacteria with resistance to various stressors and is a potential drug target. Recent Advances: We summarize recent advances in the biochemistry, structure, and physiological functions of cytochrome bd in the light of exciting new three-dimensional structures of the oxidase. The newly discovered roles of cytochrome bd in contributing to bacterial protection against hydrogen peroxide, nitric oxide, perox- ynitrite, and hydrogen sulfide are assessed. Critical Issues: Fundamental questions remain regarding the precise delineation of electron flow within this multihaem oxidase and how the extraordinarily high affinity for oxygen is accomplished, while endowing bacteria with resistance to other small ligands. Future Directions: It is clear that cytochrome bd is unique in its ability to confer resistance to toxic small molecules, a property that is significant for understanding the propensity of pathogens to possess this oxidase.
    [Show full text]
  • Multi-Omic Understanding of the Evolution of Xenobiotic Tolerance in Bacterial Isolates and Communities
    Washington University in St. Louis Washington University Open Scholarship Arts & Sciences Electronic Theses and Dissertations Arts & Sciences Summer 8-15-2019 Multi-omic Understanding of the Evolution of Xenobiotic Tolerance in Bacterial Isolates and Communities Tayte Paul Campbell Washington University in St. Louis Follow this and additional works at: https://openscholarship.wustl.edu/art_sci_etds Part of the Bioinformatics Commons, Biology Commons, and the Microbiology Commons Recommended Citation Campbell, Tayte Paul, "Multi-omic Understanding of the Evolution of Xenobiotic Tolerance in Bacterial Isolates and Communities" (2019). Arts & Sciences Electronic Theses and Dissertations. 1888. https://openscholarship.wustl.edu/art_sci_etds/1888 This Dissertation is brought to you for free and open access by the Arts & Sciences at Washington University Open Scholarship. It has been accepted for inclusion in Arts & Sciences Electronic Theses and Dissertations by an authorized administrator of Washington University Open Scholarship. For more information, please contact [email protected]. WASHINGTON UNIVERSITY IN ST. LOUIS Division of Biology and Biomedical Sciences Plant and Microbial Biosciences Dissertation Examination Committee: Gautam Dantas, Chair Arpita Bose Andrew Kau Audrey Odom-John Himadri Pakrasi Fuzhong Zhang Multi-omic Understanding of the Evolution of Xenobiotic Tolerance in Bacterial Isolates and Communities by Tayte P. Campbell A dissertation presented to The Graduate School of Washington University in partial fulfillment
    [Show full text]
  • In-Vitro Synthesis and Reconstitution of Cytochrome Bo3 Ubiquinol Oxidase in Artificial Membranes
    In-vitro Synthesis and Reconstitution of Cytochrome bo3 Ubiquinol Oxidase in Artificial Membranes Dissertation Zur Erlangung des Grades Doktor der Naturwissenschaften Am Fachbereich Biologie Der Johannes Gutenberg-Universität Mainz Ahu ARSLAN YILDIZ geboren am 20.12.1980 in Turkey Mainz, 2010 Dekan: Prof. Dr. Erwin Schmidt 1. Berichterstatter: Prof. Dr. Eva K. Sinner 2. Berichterstatter: Prof. Dr. Harald Paulsen 3. Prüfer: Prof. Dr. Wolfgang Knoll 4. Prüfer: Prof. Dr. Elmar Jaenicke Tag der mündlichen Prüfung: 29.01.2010 Abbreviations ATP Adenosine triphosphate PD Parkinson’s Disease Cyt-bo3 Cytochrome bo3 ubiquinol oxidase NADH Nicotinamide adenine dinucleotide UQ Ubiquinone UQH2 Ubiquinol Cyt-c Cytochrome c E.coli Escherichia coli Heme Heme or Haem molecule Cyt-bd Cytochrome bd ubiquinol oxidase cyo Cytochrome encoding operon CuA Copper site in subunit II CuB Copper of binuclear site CFPS Cell-free protein synthesis tRNA Transfer RNA GTP Guanidine triphosphate BLMs Black lipid membranes sBLM Supported bilayer lipid membrane tBLM Tethered lipid bilayer membrane SPR Surface Plasmon Resonance Spectroscopy SPFS Surface Plasmon Enhanced Fluorescence Spectroscopy θc Critical angle θm Minimum angle TIR Total internal reflection εd Dielectric constant n Refractive index CA Contact angle pJRHisA Subunit II histidine tagged enzyme with natural promoter pRCO3 Fused subunit II-I-III enzyme with natural promoter pETcyo Subunit II histidine tagged enzyme with T7 promoter LB Luria Bertani media ii KOH Potassium hydroxide PES Polyethylene sulfonate
    [Show full text]
  • Metabolic and Regulatory Rearrangements Underlying Glycerol Metabolism in 3 Pseudomonas Putida KT2440 4 5 by 6 7 Pablo I
    1 1 2 Metabolic and regulatory rearrangements underlying glycerol metabolism in 3 Pseudomonas putida KT2440 4 5 by 6 7 Pablo I. Nikel, Juhyun Kim, and Víctor de Lorenzo* 8 9 Systems and Synthetic Biology Program, Centro Nacional de Biotecnología (CNB-CSIC), 10 Madrid 28049, Spain 11 12 13 14 Running Title: Glycerol metabolism in P. putida 15 Keywords: Glycerol, Pseudomonas putida, central metabolism, Entner-Doudoroff pathway, redox 16 homeostasis, transcriptional regulation 17 18 19 20 _________________________________________________________________________________ 21 22 * Corresponding author V. de Lorenzo 23 Systems and Synthetic Biology Program 24 Centro Nacional de Biotecnología (CNB-CSIC) 25 Darwin 3, Campus de Cantoblanco 26 Madrid 28049, Spain 27 Phone: +34 91 585 4536 28 Fax: +34 91 585 4506 29 E-mail: [email protected] 30 2 1 SUMMARY 2 3 While the natural niches of the soil bacterium Pseudomonas putida are unlikely to include significant 4 amounts of free glycerol as a growth substrate, this bacterium is genetically equipped with the functions 5 required for its metabolism. We have resorted to deep sequencing of the transcripts in glycerol-grown 6 P. putida KT2440 cells to gain an insight into the biochemical and regulatory components involved in 7 the shift between customary C sources (e.g., glucose or succinate) to the polyol. Transcriptomic results 8 were contrasted with key enzymatic activities under the same culture conditions. Cognate expression 9 profiles revealed that genes encoding enzymes of the Entner-Doudoroff route and other catabolic 10 pathways, e.g., the gluconate and 2-ketogluconate loops, were significantly down-regulated on glycerol.
    [Show full text]
  • Differential Effects of Epinephrine, Norepinephrine, and Indole
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Texas A&M University INFECTION AND IMMUNITY, Sept. 2007, p. 4597–4607 Vol. 75, No. 9 0019-9567/07/$08.00ϩ0 doi:10.1128/IAI.00630-07 Copyright © 2007, American Society for Microbiology. All Rights Reserved. Differential Effects of Epinephrine, Norepinephrine, and Indole on Escherichia coli O157:H7 Chemotaxis, Colonization, and Gene Expressionᰔ Tarun Bansal, Derek Englert, Jintae Lee, Manjunath Hegde, Thomas K. Wood, and Arul Jayaraman* Artie McFerrin Department of Chemical Engineering, Texas A&M University, College Station, Texas 77843-3122 Received 3 May 2007/Returned for modification 4 June 2007/Accepted 13 June 2007 During infection in the gastrointestinal tract, enterohemorrhagic Escherichia coli (EHEC) O157:H7 is exposed to a wide range of signaling molecules, including the eukaryotic hormones epinephrine and norepi- nephrine, and bacterial signal molecules such as indole. Since these signaling molecules have been shown to be involved in the regulation of phenotypes such as motility and virulence that are crucial for EHEC infections, we hypothesized that these molecules also govern the initial recognition of the large intestine environment and attachment to the host cell surface. Here, we report that, compared to indole, epinephrine and norepinephrine exert divergent effects on EHEC chemotaxis, motility, biofilm formation, gene expression, and colonization of HeLa cells. Using a novel two-fluorophore chemotaxis assay, it was found that EHEC is attracted to epineph- rine and norepinephrine while it is repelled by indole. In addition, epinephrine and norepinephrine also increased EHEC motility and biofilm formation while indole attenuated these phenotypes.
    [Show full text]
  • Davidge JBC Supplementary.Pdf
    promoting access to White Rose research papers Universities of Leeds, Sheffield and York http://eprints.whiterose.ac.uk/ White Rose Research Online URL for this paper: http://eprints.whiterose.ac.uk/7923/ (includes links to Main Article, Supplementary Material and Figures) Published paper Davidge, K.S., Sanguinetti, G., Yee, C.H., Cox, A.G., McLeod, C.W., Monk, C.E., Mann, B.E., Motterlini, R. and Poole, R.K. (2009) Carbon monoxide-releasing antibacterial molecules target respiration and global transcriptional regulators. Journal of Biological Chemistry, 284 (7). pp. 4516-4524. http://dx.doi.org/10.1074/jbc.M808210200 Supplementary Material White Rose Research Online [email protected] Supplementary Material Carbon monoxide-releasing antibacterial molecules target respiration and global transcriptional regulators Kelly S Davidge, Guido Sanguinetti, Chu Hoi Yee, Alan G Cox, Cameron W McLeod, Claire E Monk, Brian E Mann, Roberto Motterlini and Robert K Poole Contents Page Number Supplementary Figure S1 3 Inhibition by CORM-3 of E. coli cultures grown in defined medium anaerobically and aerobically Supplementary Figure S2 4 Viability assays showing survival of anaerobically and aerobically E. coli in defined growth medium Supplementary Figure S3 5 Reaction of terminal oxidases in vivo on addition of RuCl2(DMSO)4 to intact cells in a dual-wavelength spectrophotometer Supplementary Figure S4 6 CORM-3 generates carbonmonoxycytochrome bd in vivo and depresses synthesis of cytochrome bo' Supplementary Figure S5 7 Expression of spy-lacZ activity
    [Show full text]
  • Letters to Nature
    letters to nature Received 7 July; accepted 21 September 1998. 26. Tronrud, D. E. Conjugate-direction minimization: an improved method for the re®nement of macromolecules. Acta Crystallogr. A 48, 912±916 (1992). 1. Dalbey, R. E., Lively, M. O., Bron, S. & van Dijl, J. M. The chemistry and enzymology of the type 1 27. Wolfe, P. B., Wickner, W. & Goodman, J. M. Sequence of the leader peptidase gene of Escherichia coli signal peptidases. Protein Sci. 6, 1129±1138 (1997). and the orientation of leader peptidase in the bacterial envelope. J. Biol. Chem. 258, 12073±12080 2. Kuo, D. W. et al. Escherichia coli leader peptidase: production of an active form lacking a requirement (1983). for detergent and development of peptide substrates. Arch. Biochem. Biophys. 303, 274±280 (1993). 28. Kraulis, P.G. Molscript: a program to produce both detailed and schematic plots of protein structures. 3. Tschantz, W. R. et al. Characterization of a soluble, catalytically active form of Escherichia coli leader J. Appl. Crystallogr. 24, 946±950 (1991). peptidase: requirement of detergent or phospholipid for optimal activity. Biochemistry 34, 3935±3941 29. Nicholls, A., Sharp, K. A. & Honig, B. Protein folding and association: insights from the interfacial and (1995). the thermodynamic properties of hydrocarbons. Proteins Struct. Funct. Genet. 11, 281±296 (1991). 4. Allsop, A. E. et al.inAnti-Infectives, Recent Advances in Chemistry and Structure-Activity Relationships 30. Meritt, E. A. & Bacon, D. J. Raster3D: photorealistic molecular graphics. Methods Enzymol. 277, 505± (eds Bently, P. H. & O'Hanlon, P. J.) 61±72 (R. Soc. Chem., Cambridge, 1997).
    [Show full text]
  • Structure of the Trypanosome Cyanide-Insensitive Alternative Oxidase
    Structure of the trypanosome cyanide-insensitive alternative oxidase Tomoo Shibaa,1,2, Yasutoshi Kidoa,1,3, Kimitoshi Sakamotoa,4, Daniel Ken Inaokaa, Chiaki Tsugea, Ryoko Tatsumia, Gen Takahashib, Emmanuel Oluwadare Baloguna,b,c, Takeshi Narad, Takashi Aokid, Teruki Honmae, Akiko Tanakae, Masayuki Inouef, Shigeru Matsuokaf, Hiroyuki Saimotog, Anthony L. Mooreh, Shigeharu Haradab,5, and Kiyoshi Kitaa,5 aDepartment of Biomedical Chemistry, Graduate School of Medicine, and fGraduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033, Japan; bDepartment of Applied Biology, Graduate School of Science and Technology, Kyoto Institute of Technology, Kyoto 606-8585, Japan; cDepartment of Biochemistry, Ahmadu Bello University, Zaria 2222, Nigeria; dDepartment of Molecular and Cellular Parasitology, Juntendo University School of Medicine, Tokyo 113-8421, Japan; eSystems and Structural Biology Center, RIKEN, Tsurumi, Yokohama 230-0045, Japan; gDepartment of Chemistry and Biotechnology, Graduate School of Engineering, Tottori University, Tottori 680-8552, Japan; and hBiochemistry and Molecular Biology, School of Life Sciences, University of Sussex, Brighton BN1 9QG, United Kingdom Edited† by John E. Walker, Medical Research Council Mitochondrial Biology Unit, Cambridge, United Kingdom, and approved February 11, 2013 (received for review October 23, 2012) In addition to haem copper oxidases, all higher plants, some algae, mammalian host in the bloodstream, both the cytochrome re- yeasts, molds, metazoans, and pathogenic microorganisms such as spiratory pathway and oxidative phosphorylation disappear and Trypanosoma brucei contain an additional terminal oxidase, the are replaced by the trypanosomal alternative oxidase (TAO), cyanide-insensitive alternative oxidase (AOX). AOX is a diiron car- which functions as the sole terminal oxidase to reoxidize NADH boxylate protein that catalyzes the four-electron reduction of accumulated during glycolysis (5).
    [Show full text]
  • Cyddc-Mediated Reductant Export in Escherichia Coli Controls the Transcriptional Wiring of Energy Metabolism and Combats Nitrosative Stress Louise V
    Biochem. J. (2016) 473, 693–701 doi:10.1042/BJ20150536 693 CydDC-mediated reductant export in Escherichia coli controls the transcriptional wiring of energy metabolism and combats nitrosative stress Louise V. Holyoake*, Stuart Hunt†, Guido Sanguinetti‡, Gregory M. Cook§, Mark J. Howard*, Michelle L. Rowe*, Robert K. Poole† and Mark Shepherd*1 *School of Biosciences, University of Kent, Canterbury CT2 7NJ, U.K. †Department of Molecular Biology and Biotechnology, The University of Sheffield, Sheffield S10 2TN, U.K. ‡School of Informatics, The University of Edinburgh, Informatics Forum, 10 Crichton Street, Edinburgh EH8 9AB, U.K. §Department of Microbiology and Immunology, University of Otago, P.O. Box 56, 720 Cumberland Street, Dunedin 9054, New Zealand The glutathione/cysteine exporter CydDC maintains redox CydDC-mediated reductant export promotes protein misfolding, Downloaded from http://portlandpress.com/biochemj/article-pdf/473/6/693/686013/bj4730693.pdf by guest on 02 October 2021 balance in Escherichia coli.AcydD mutant strain was adaptations to energy metabolism and sensitivity to NO. The used to probe the influence of CydDC upon reduced thiol addition of both glutathione and cysteine to the medium was found export, gene expression, metabolic perturbations, intracellular to complement the loss of bd-type cytochrome synthesis in a cydD pH homoeostasis and tolerance to nitric oxide (NO). Loss strain (a key component of the pleiotropic cydDC phenotype), of CydDC was found to decrease extracytoplasmic thiol providing the first direct evidence that CydDC substrates are levels, whereas overexpression diminished the cytoplasmic able to restore the correct assembly of this respiratory oxidase. thiol content. Transcriptomic analysis revealed a dramatic These data provide an insight into the metabolic flexibility of E.
    [Show full text]
  • Product Sheet Info
    Master Clone List for NR-19279 ® Vibrio cholerae Gateway Clone Set, Recombinant in Escherichia coli, Plates 1-46 Catalog No. NR-19279 Table 1: Vibrio cholerae Gateway® Clones, Plate 1 (NR-19679) Clone ID Well ORF Locus ID Symbol Product Accession Position Length Number 174071 A02 367 VC2271 ribD riboflavin-specific deaminase NP_231902.1 174346 A03 336 VC1877 lpxK tetraacyldisaccharide 4`-kinase NP_231511.1 174354 A04 342 VC0953 holA DNA polymerase III, delta subunit NP_230600.1 174115 A05 388 VC2085 sucC succinyl-CoA synthase, beta subunit NP_231717.1 174310 A06 506 VC2400 murC UDP-N-acetylmuramate--alanine ligase NP_232030.1 174523 A07 132 VC0644 rbfA ribosome-binding factor A NP_230293.2 174632 A08 322 VC0681 ribF riboflavin kinase-FMN adenylyltransferase NP_230330.1 174930 A09 433 VC0720 phoR histidine protein kinase PhoR NP_230369.1 174953 A10 206 VC1178 conserved hypothetical protein NP_230823.1 174976 A11 213 VC2358 hypothetical protein NP_231988.1 174898 A12 369 VC0154 trmA tRNA (uracil-5-)-methyltransferase NP_229811.1 174059 B01 73 VC2098 hypothetical protein NP_231730.1 174075 B02 82 VC0561 rpsP ribosomal protein S16 NP_230212.1 174087 B03 378 VC1843 cydB-1 cytochrome d ubiquinol oxidase, subunit II NP_231477.1 174099 B04 383 VC1798 eha eha protein NP_231433.1 174294 B05 494 VC0763 GTP-binding protein NP_230412.1 174311 B06 314 VC2183 prsA ribose-phosphate pyrophosphokinase NP_231814.1 174603 B07 108 VC0675 thyA thymidylate synthase NP_230324.1 174474 B08 466 VC1297 asnS asparaginyl-tRNA synthetase NP_230942.2 174933 B09 198
    [Show full text]
  • The Thuggacins, Novel Antibacterial Macrolides from Sorangium
    J. Antibiot. 60(12): 733–738, 2007 THE JOURNAL OF ORIGINAL ARTICLE ANTIBIOTICS The Thuggacins, Novel Antibacterial Macrolides from Sorangium cellulosum Acting against Selected Gram- positive Bacteria Production, Antimicrobial Activity and Mechanism of Action Herbert Irschik, Hans Reichenbach, Gerhard Höfle, Rolf Jansen Received: August 16, 2007 / Accepted: November 28, 2007 © Japan Antibiotics Research Association Abstract In our screening program we found an and the structures were elucidated [2] (Fig. 1). activity against some Gram-positive bacteria, including This report gives some data on the production mycobacteria in the culture supernatant of Sorangium and the biological activities of thuggacins A, B and C. cellulosum strain So ce895. The antibiotic responsible for Thuggacin A acts on the respiration of some bacteria. this activity was isolated and named thuggacin. Initial Electron transport chains in microorganisms with their studies towards the mechanism of action showed that different enzyme complexes are described in [3]. NADH thuggacin A inhibits a late step of the respiratory chain of driven electron flow begins with the oxidation of some bacteria. NADH and the reduction of ubiquinone by NADH- oxidase. Then in several bacteria, e.g. Micrococcus luteus Keywords Sorangium cellulosum, antibiotics, macro- and Mycobacterium phlei, electrons are transferred to lides, respiration, thuggacin cytochrome c by an enzyme complex containing cytochromes bc1. The last step is the reduction of oxygen by the cytochrome c oxidase (terminal oxidase), containing the Introduction cytochromes aa3. Other bacteria like Escherichia coli oxidize the ubiquinone by an ubiquinol oxidase. In most In the last few years, we isolated many strains of the cases bacteria have more than one terminal oxidase with order myxobacteria from soil samples collected from different cytochromes.
    [Show full text]
  • The Role and Regulation of Quinol Oxidase in Rhizobium Etli Zachary Ryan Lunak Marquette University
    Marquette University e-Publications@Marquette Dissertations (2009 -) Dissertations, Theses, and Professional Projects The Role and Regulation of Quinol Oxidase in Rhizobium Etli Zachary Ryan Lunak Marquette University Recommended Citation Lunak, Zachary Ryan, "The Role and Regulation of Quinol Oxidase in Rhizobium Etli" (2015). Dissertations (2009 -). Paper 504. http://epublications.marquette.edu/dissertations_mu/504 THE ROLE AND REGULATION OF QUINOL OXIDASE IN RHIZOBIUM ETLI by Zachary Lunak, B.S., M.S. A Dissertation Submitted to the Faculty of the Graduate School, Marquette University, in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy Milwaukee Wisconsin May 2015 ABSTRACT THE ROLE AND REGULATION OF QUINOL OXIDASE IN RHIZOBIUM ETLI Zachary Lunak, B.S., M.S. Marquette University, 2015 Aerobic bacteria typically respire by delivering electrons to oxygen via cytochrome c oxidases. In addition, many bacteria can respire by transferring electrons to oxygen via quinol oxidases. Exactly why bacteria contain quinol oxidases when they already have cytochrome c oxidases is unclear. In particular, the Cyo quinol oxidase, encoded by cyoABCD, is widespread among the dominant bacterial groups but its specific role is unknown. Rhizobium etli CFN42 provides an aerobic respiratory model in which Cyo can be directly compared to cytochrome c oxidases within the same organism. Mutants of the terminal oxidases in R. etli were constructed and their ability to grow in different physiological conditions was examined. The cyo mutant had noticable growth defects in low oxygen, low pH and low iron conditions. Furthermore, the cyo gene in the wild type was significantly up-regulated in these conditions. Conversely, in slow-growth conditions, such as stationary growth or growth in certain carbon sources, cyo was significantly down-regulated.
    [Show full text]