HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Nguyen, H.-H. T. Articles by Chan, S. I. Search for Related Content PubMed PubMed Citation Articles by Nguyen, H.-H. T. Articles by Chan, S. I. Social Bookmarking                      What's this?

J Biol Chem, Vol. 273, Issue 14, 7957-7966, April 3, 1998

The Particulate Methane Monooxygenase from Methylococcus capsulatus (Bath) Is a Novel Copper-containing Three-subunit Enzyme
ISOLATION AND CHARACTERIZATION

From the Arthur Amos Noyes Laboratory of Chemical Physics, California Institute of Technology, Pasadena, California 91125

The particulate methane monooxygenase (pMMO) is known to be very difficult to study mainly due to its unusual activity instability in vitro. By cultivating Methylococcus capsulatus (Bath) under methane stress conditions and high copper levels in the growth medium, membranes highly enriched in the pMMO with exceptionally stable activity can be isolated from these cells. Purified and active pMMO can be subsequently obtained from these membrane preparations using protocols in which an excess of reductants and anaerobic conditions were maintained during membrane solubilization by dodecyl -D-maltoside and purification by chromatography. The pMMO was found to be the major constituent in these membranes, constituting 60-80% of total membrane proteins. The dominant species of the pMMO was found to consist of three subunits, , , and , with an apparent molecular mass of 45, 26, and 23 kDa, respectively. A second species of the pMMO, a proteolytically processed version of the enzyme, was found to be composed of three subunits, ', , and , with an apparent molecular mass of 35, 26, and 23 kDa, respectively. The  and ' subunits from these two forms of the pMMO contain identical N-terminal sequences. The  subunit, however, exhibits variation in its N-terminal sequence. The pMMO is a copper-containing protein only and shows a requirement for Cu(I) ions. Approximately 12-15 Cu ions per 94-kDa monomeric unit were observed. The pMMO is sensitive to dioxygen tension. On the basis of dioxygen sensitivity, three kinetically distinct forms of the enzyme can be distinguished. A slow but air-stable form, which is converted into a "pulsed" state upon direct exposure to atmospheric oxygen pressure, is considered as type I pMMO. This form was the subject of our pMMO isolation effort. Other forms (types II and III) are deactivated to various extents upon exposure to atmospheric dioxygen pressure. Under inactivating conditions, these unstable forms release protons to the buffer (~10 H⁺/94-kDa monomeric unit) and eventually become completely inactive.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				H. Ali, J. Scanlan, M. G. Dumont, and J. C. Murrell Duplication of the mmoX gene in Methylosinus sporium: cloning, sequencing and mutational analysis. Microbiology, October 1, 2006; 152(Pt 10): 2931 - 2942. [Abstract] [Full Text] [PDF]

				D. W. Choi, W. E. Antholine, Y. S. Do, J. D. Semrau, C. J. Kisting, R. C. Kunz, D. Campbell, V. Rao, S. C. Hartsel, and A. A. DiSpirito Effect of methanobactin on the activity and electron paramagnetic resonance spectra of the membrane-associated methane monooxygenase in Methylococcus capsulatus Bath Microbiology, October 1, 2005; 151(10): 3417 - 3426. [Abstract] [Full Text] [PDF]

				H. Dalton The Leeuwenhoek Lecture 2000 The natural and unnatural history of methane-oxidizing bacteria Phil Trans R Soc B, June 29, 2005; 360(1458): 1207 - 1222. [Abstract] [Full Text] [PDF]

				D. P. Erwin, I. K. Erickson, M. E. Delwiche, F. S. Colwell, J. L. Strap, and R. L. Crawford Diversity of Oxygenase Genes from Methane- and Ammonia-Oxidizing Bacteria in the Eastern Snake River Plain Aquifer Appl. Envir. Microbiol., April 1, 2005; 71(4): 2016 - 2025. [Abstract] [Full Text] [PDF]

				P. Ricke, C. Erkel, M. Kube, R. Reinhardt, and W. Liesack Comparative Analysis of the Conventional and Novel pmo (Particulate Methane Monooxygenase) Operons from Methylocystis Strain SC2 Appl. Envir. Microbiol., May 1, 2004; 70(5): 3055 - 3063. [Abstract] [Full Text] [PDF]

				A. K. Shiemke, D. J. Arp, and L. A. Sayavedra-Soto Inhibition of Membrane-Bound Methane Monooxygenase and Ammonia Monooxygenase by Diphenyliodonium: Implications for Electron Transfer J. Bacteriol., February 15, 2004; 186(4): 928 - 937. [Abstract] [Full Text] [PDF]

				J. L. Blazyk and S. J. Lippard Domain Engineering of the Reductase Component of Soluble Methane Monooxygenase from Methylococcus capsulatus (Bath) J. Biol. Chem., February 13, 2004; 279(7): 5630 - 5640. [Abstract] [Full Text] [PDF]

				S. S.-F. Yu, L.-Y. Wu, K. H.-C. Chen, W.-I Luo, D.-S. Huang, and S. I. Chan The Stereospecific Hydroxylation of [2,2-2H2]Butane and Chiral Dideuteriobutanes by the Particulate Methane Monooxygenase from Methylococcus capsulatus (Bath) J. Biol. Chem., October 17, 2003; 278(42): 40658 - 40669. [Abstract] [Full Text] [PDF]

				S. S.-F. Yu, K. H.-C. Chen, M. Y.-H. Tseng, Y.-S. Wang, C.-F. Tseng, Y.-J. Chen, D.-S. Huang, and S. I. Chan Production of High-Quality Particulate Methane Monooxygenase in High Yields from Methylococcus capsulatus (Bath) with a Hollow-Fiber Membrane Bioreactor J. Bacteriol., October 15, 2003; 185(20): 5915 - 5924. [Abstract] [Full Text] [PDF]

				D.-W. Choi, R. C. Kunz, E. S. Boyd, J. D. Semrau, W. E. Antholine, J.-I. Han, J. A. Zahn, J. M. Boyd, A. M. de la Mora, and A. A. DiSpirito The Membrane-Associated Methane Monooxygenase (pMMO) and pMMO-NADH:Quinone Oxidoreductase Complex from Methylococcus capsulatus Bath J. Bacteriol., October 1, 2003; 185(19): 5755 - 5764. [Abstract] [Full Text] [PDF]

				R. L. Lieberman, D. B. Shrestha, P. E. Doan, B. M. Hoffman, T. L. Stemmler, and A. C. Rosenzweig Bioinorganic Chemistry Special Feature: Purified particulate methane monooxygenase from Methylococcus capsulatus (Bath) is a dimer with both mononuclear copper and a copper-containing cluster PNAS, April 1, 2003; 100(7): 3820 - 3825. [Abstract] [Full Text] [PDF]

				P. F. Dunfield and R. Conrad Starvation Alters the Apparent Half-Saturation Constant for Methane in the Type II Methanotroph Methylocystis Strain LR1 Appl. Envir. Microbiol., September 1, 2000; 66(9): 4136 - 4138. [Abstract] [Full Text]

				J. D. Morton, K. F. Hayes, and J. D. Semrau Effect of Copper Speciation on Whole-Cell Soluble Methane Monooxygenase Activity in Methylosinus trichosporium OB3b Appl. Envir. Microbiol., April 1, 2000; 66(4): 1730 - 1733. [Abstract] [Full Text]

				B. Gilbert, I. R. McDonald, R. Finch, G. P. Stafford, A. K. Nielsen, and J. C. Murrell Molecular Analysis of the pmo (Particulate Methane Monooxygenase) Operons from Two Type II Methanotrophs Appl. Envir. Microbiol., March 1, 2000; 66(3): 966 - 975. [Abstract] [Full Text]

				S. Grosse, L. Laramee, K.-D. Wendlandt, I. R. McDonald, C. B. Miguez, and H.-P. Kleber Purification and Characterization of the Soluble Methane Monooxygenase of the Type II Methanotrophic Bacterium Methylocystis sp. Strain WI 14 Appl. Envir. Microbiol., September 1, 1999; 65(9): 3929 - 3935. [Abstract] [Full Text]

				A. A. DiSpirito, J. A. Zahn, D. W. Graham, H. J. Kim, C. K. Larive, T. S. Derrick, C. D. Cox, and A. Taylor Copper-Binding Compounds from Methylosinus trichosporium OB3b J. Bacteriol., July 15, 1998; 180(14): 3606 - 3613. [Abstract] [Full Text]