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Cytochrome aa 3 from Methylococcus capsulatus (Bath)

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Abstract

A cytochrome aa 3-type oxidase was isolated with and without a c-type cytochrome (cytochrome c-557) from Methylococcus capsulatus Bath by ion-exchange and hydrophobic chromatography in the presence of Triton X-100. Although cytochrome c-557 was not a constitutive component of the terminal oxidase, the cytochrome c ascorbate-TMPD oxidase activity of the enzyme decreased dramatically when the ratio of cytochrome c-557 to heme a dropped below 1:3. On denaturing gels, the purified enzyme dissociated into three subunits with molecular weights of 46,000, 28,000 and 20,000. The enzyme contains two heme groups (a and a 3), absorption maximum at 422 nm in the resting state, at 445 and 601 nm in the dithionite reduced form and at 434 and 598 nm in the dithionite reduced plus CO form. Denaturing gels of the cytochrome aa 3-cytochrome c-557 complex showed the polypeptides associated with cytochrome aa 3 plus a heme c-staining subunit with a molecular weight of 37,000. The complex contains approximately two heme a, one heme c, absorption maximum at 420 nm in the resting state and at 421, 445, 522, 557 and 601 nm in the dithionite reduced form. The specific activity of the purified enzyme was 130 mol O2/min · mol heme a compared to 753 mol O2/min · mol heme a when isolated with cytochrome c-557.

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Abbreviations

MMO:

methan monooxygenase

sMMO:

soluble methane monooxygenase

pMMO:

particulate methane monooxygenase

TMPD:

N,N,N′,N′-tetramethyl-p-phenylenediamine dihydrochloride

Na2EDTA:

disodium ethylenediamine-tetraacetic acid

References

  • Ankent'eva NF, Govozdev RI (1988) Purification and physicochemical properties of methane monooxygenase from membrane structures of Methylococcus capsulatus. Microbiology 53: 93–96.

    Google Scholar 

  • Anraku Y (1988) Bacterial electron transport chains. Ann Rev Biochem 57: 101–132.

    Google Scholar 

  • Capaldi RA, Vanderkooi G (1972) The low polarity of many membrane proteins. Proc Natl Acad Sci USA 69: 930–932.

    Google Scholar 

  • Chan SI, Nguyen HT, Shiemke AK, Lidstrom ME (1993) Biochemical and biophysical studies toward characterization of the membrane-associated methane monooxygenase. In: Murrell JC, Kelly DP (eds) Microbial growth on C1 compounds Intercept Ltd, Hampshire, UK.

    Google Scholar 

  • Dalton H, Whittenbury R (1976) The acetylene reduction technique as an assay for nitrogen fixation activity in the methane oxidizing bacterium Methylococcus capsulatus strain Bath. Arch Microbiol 109: 147–151.

    Google Scholar 

  • Dalton H, Prior SD, Stanley SH (1984) Regulation and control of methane monooxygenase. In: Crawford RL, Hanson RS (eds) Microbial growth on c 1 compounds. American Society for Microbiology Press, Washington DC, pp 75–82.

    Google Scholar 

  • Day DJ, Anthony C (1990) Methanol dehydrogenase from Methylobacterium extorquens AM1. Methods Enzymol 188: 210–216.

    Google Scholar 

  • DiSpirito AA (1990) Soluble cytochromes c from Methylomonas A4. Methods Enzymol 188: 289–297.

    Google Scholar 

  • DiSpirito AA, Lipscomb JD, Hooper AB (1986) Cytochrome aa 3 from Nitrosomonas europaea. J Biol Chem 261: 17048–17056.

    Google Scholar 

  • DiSpirito AA, Lipscomb JD, Lidstrom ME (1990) Soluble cytochromes from the marine methanotroph Methylomonas sp. strain A4. J Bacteriol 172: 5360–5367.

    Google Scholar 

  • DiSpirito AA, Gulledge JA, Shiemke AK, Murrell JC, Lidstrom ME, Krema CL (1992) Trichloroethylene oxidation by the membrane-associated methane monooxygenase in type I, type II and type X methanotrophs. Biodegradation 2: 151–164.

    Google Scholar 

  • Fee JA, Choc MG, Findling KL, Lorance R, Yoshida T (1980) Properties of a copper-containing cytochrome c 1 aa 3 complex: a terminal oxidase of the extreme thermophile Thermus thermophilus HB8. Proc Natl Acad Sci USA 77: 147–161.

    Google Scholar 

  • Ferguson-Miller S, Brautigan DL, Margoliash E (1976) Correlation of the kinetics of electron transfer activity of various eukaryotic cytochromes c with binding to mitochondrial cytochrome c oxidase. J Biol Chem 251: 1104–1115.

    Google Scholar 

  • Fuhrhop J-H (1975) Laboratory method in porphyrin and metalloporphyrin research. Elsevier Scientific Publishers, New York.

    Google Scholar 

  • Gennis RB (1991) Some recent advances relating to prokaryotic cytochrome c reductases and cytochrome c oxidases. Biochim Biophys Acta 1058: 21–24.

    Google Scholar 

  • Hon-nami K, Oshima T (1980) Cytochrome oxidase from an extreme thermophile, Thermus thermophilus HB8. Biochem Biophys Res Commun 92: 1023–1029.

    Google Scholar 

  • Kadenbach B, Jarausch J, Hartmann R, Merle P (1983) Separation of mammalian cytochrome c oxidase into 13 polypeptides by a sodium dodecyl sulfate-gel electrophoretic procedure. Anal Biochem 129: 517–521.

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 27: 680–685.

    Google Scholar 

  • Lidstrom ME (1988) Isolation and characterization of marine methanotrophs. Antonie van Leeuvenhoek. J Microbiol Serol 54: 189–199.

    Google Scholar 

  • Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275.

    Google Scholar 

  • Ludwig B (1987) Cytochrome c oxidase in prokaryotes. FEMS Microbiol Rev 46: 41–56.

    Google Scholar 

  • Ludwig B, Schatz G (1980) A two-subunit cytochrome c oxidase (cytochrome aa 3) from Paracoccus denitrificans. Proc Natl Acad Sci USA 77: 196–200.

    Google Scholar 

  • Ludwig B, Gibson QH (1981) Reaction of oxygen with cytochrome c oxidase from Paracoccus denitrificans. J Biol Chem 256: 10092–10098.

    Google Scholar 

  • McDonnel A, Staehelin LA (1981) Detection of cytochrome f. a c-class cytchrome, with diaminobenzidine in polyacrylamide gels. Anal Biochem 117: 40–44.

    Google Scholar 

  • Merle P, Kadenbach B (1980) The subunit composition of mammalian cytochrome c oxidase. Eur J Biochem 105: 499–507.

    Google Scholar 

  • Naqui A, Powers L, Lundee M, Constantinescu A, Chance B (1988) On the environment of zinc in beef heart cytochrome c oxidase: an X-ray absorption study. J Biol Chem 263: 12342–12345.

    Google Scholar 

  • Öblad M, Selin E, Malmström, Strid L, Aasa R, Malmström BG (1989) Analytical characterization of cytochrome oxidase preparations with regard to metal and phospholipid contents, peptide composition and catalytic activity. Biochim Biophys Acta 975: 267–270.

    Google Scholar 

  • Pan L-P, He Q, Chan SI (1991) The nature of zinc in cytochrome c oxidase. J Biol Chem 266: 19109–19112.

    Google Scholar 

  • Poynton RO, Schatz G (1975) Cytochrome c oxidized from Bakers'yeast. III. Physical characterization of isolated subunits and chemical evidence for two different classes of polypeptides. J Biol Chem 250: 752–761.

    Google Scholar 

  • Prior SD, Dalton H (1985a) Acetylene as a suicide substrate and active site probe for the membrane methane monooxygenase from Methylococcus capsulatus (Bath). FEMS Microbiol Lett 29: 105–109.

    Google Scholar 

  • Prior SD, Dalton H (1985b) The effect of copper ions on membrane content and methane monooxygenase activity in methanolgrown cells of Methylococcus capsulatus (Bath). J Gen Microbiol 131: 155–163.

    Google Scholar 

  • Robinson J, Cooper JM (1970) Method of determining oxygen concentrations in biological media, suitable for calibration of oxygen electrode. Anal Biochem 33: 390–399.

    Google Scholar 

  • Scott D, Brannan J, Higgins IJ (1981) The effect of growth conditions on intracytoplasmic membranes and methane monooxygenase activities in Methylosinus trichosporium OB3b. J Gen Microbiol 125: 63–72.

    Google Scholar 

  • Smith DDS, Dalton H (1989) Solubilization of methane monooxygenase from Methylococcus capsulatus (Bath). Eur J Biochem 182: 667–671.

    Google Scholar 

  • Sone N, Yanagita S (1982) A cytochrome aa 3-type terminal oxidase of a thermophilic bacterium purification, properties and proton pumping. Biochem Biophys Acta 682: 216–226.

    Google Scholar 

  • Stanley SH, Prior SD, Leak DJ, Dalton H (1983) Copper stress underlies the fundamental change in intracellular location of methane mono-oxygenase in methane-oxidizing organisms: studies in batch and continuous cultures. Biotechnol Lett 5: 487–492.

    Google Scholar 

  • Stirling DI, Dalton H (1977) Effect of metal-binding agents and other compounds on methane oxidation by two strains of Methylococcus capsulatus. Arch Microbiol 114: 71–76.

    Google Scholar 

  • Stirling DI, Colby J, Dalton H (1979) A comparison of the substrate and electron-donor specificity of the methane mono-oxygenase from three strains of methane-oxidizing bacteria. Biochem J 177: 362–364.

    Google Scholar 

  • Tonge GM, Harrison DEF, Higgins IJ (1977) Purification and properties of the soluble methane monooxygenase from Methylosinus trichosporium OB3b. Biochem J 161: 333–334.

    Google Scholar 

  • Whittenbury R, Philips KC, Wilkinson JF (1970) Enrichment, isolation and some properties of methane utilizing bacteria. J Gen Microbiol 61: 205–218.

    Google Scholar 

  • Williams JN (1964) A method for the simultaneous quantitative estimation of cytochromes a, b, c 1, and c in mitochondria. Arch Biochem Biophys 107: 537–543.

    Google Scholar 

  • Whittenbury R, Dalton H (1981) The methylotrophic bacteria. In: Starr MP, Trüper HG, Balows A, Schlegel HG (eds) The prokaryotes, vol 1. Springer, New York, pp 894–902.

    Google Scholar 

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Authors and Affiliations

  1. Department of Microbiology, Immunology and Preventive Medicine, Iowa State University, 205 Science State Building I, 50011, Ames, IA, USA

    Alan A. DiSpirito & James A. Zahn

  2. Department of Biochemistry, West Virginia University School of Medicine, 26506, Morgantown, West Virginia, USA

    Andrew K. Shiemke

  3. Department of Microbiology, University of Illinois, 61801, Urbana, IL, USA

    Sean W. Jordan

  4. Department of Biochemistry, University of Iowa, Bowen Science Building, 52242, Iowa City, IA, USA

    Cinder L. Krema

Authors
  1. Alan A. DiSpirito
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  2. Andrew K. Shiemke
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  3. Sean W. Jordan
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  4. James A. Zahn
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  5. Cinder L. Krema
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DiSpirito, A.A., Shiemke, A.K., Jordan, S.W. et al. Cytochrome aa 3 from Methylococcus capsulatus (Bath). Arch. Microbiol. 161, 258–265 (1994). https://doi.org/10.1007/BF00248702

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  • DOI: https://doi.org/10.1007/BF00248702

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