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Global proteomic analysis of the insoluble, soluble, and supernatant fractions of the psychrophilic archaeon Methanococcoides burtonii. Part II: the effect of different methylated growth substrates

Global proteomic analysis of the insoluble, soluble, and supernatant fractions of the psychrophilic archaeon Methanococcoides burtonii. Part II: the effect of different methylated growth substrates
Global proteomic analysis of the insoluble, soluble, and supernatant fractions of the psychrophilic archaeon Methanococcoides burtonii. Part II: the effect of different methylated growth substrates
Methanococcoides burtonii is a cold-adapted methanogenic archaeon from Ace Lake in Antarctica. Methanol and methylamines are the only substrates it can use for carbon and energy. We carried out quantitative proteomics using iTRAQ of M. burtonii cells grown on different substrates (methanol in defined media or trimethylamine in complex media), using techniques that enriched for secreted and membrane proteins in addition to cytoplasmic proteins. By integrating proteomic data with the complete, manually annotated genome sequence of M. burtonii, we were able to gain new insight into methylotrophic metabolism and the effects of methanol on the cell. Metabolic processing of methanol and methylamines is initiated by methyltransferases specific for each substrate, with multiple paralogs for each of the methyltransferases (similar to other members of the Methanosarcinaceae). In M. burtonii, most methyltransferases appear to have distinct roles in the metabolism of methylated substrates, although two methylamine methyltransferases appear to be nonfunctional. One set of methyltransferases for trimethylamine catabolism appears to be membrane associated, potentially providing a mechanism to directly couple trimethylamine uptake to demethylation. Important roles were highlighted for citrate synthase, glutamine synthetase, acetyl-CoA decarbonylase/synthase, and pyruvate synthase in carbon and nitrogen metabolism during growth on methanol. M. burtonii had only a marginal response to the provision of exogenous amino acids (from yeast extract), indicating that it is predisposed to the endogenous synthesis of amino acids. Growth on methanol appeared to cause oxidative stress in the cell, possibly through the formation of reactive nonoxygen species and formaldehyde, and the oxidative inactivation of corrinoid proteins, with the cell responding by elevating the synthesis of universal stress (Usp) proteins, several nucleic acid binding proteins, and a serpin. In addition, changes in levels of cell envelope proteins were linked to counteracting the disruptive solvent effects of methanol on cell membranes. This is the first global proteomic study to examine the effects of different carbon sources on the growth of an obligately methylotrophic methanogen.
1535-3893
653-663
Williams, Timothy J.
c45d1f23-12f3-4dab-bebb-e89fb7219e26
Burg, Dominic W.
a51bc2a5-54d3-40ee-ac66-4ca0bc4d9d1d
Ertan, Haluk
25a742c1-9323-4fae-820e-33e22266b60b
Raftery, Mark J.
6050a0f6-677d-468d-ac46-76663d1ac6c3
Poljak, Anne
114fe1b4-3275-4d81-8d9c-ff523cd18254
Guilhaus, Michael
f2e34a05-c65c-41cc-9f30-268d41d5d3e7
Cavicchioli, Ricardo
95c04c14-6cdd-4f0d-afc6-32bdbfa53ad9
Williams, Timothy J.
c45d1f23-12f3-4dab-bebb-e89fb7219e26
Burg, Dominic W.
a51bc2a5-54d3-40ee-ac66-4ca0bc4d9d1d
Ertan, Haluk
25a742c1-9323-4fae-820e-33e22266b60b
Raftery, Mark J.
6050a0f6-677d-468d-ac46-76663d1ac6c3
Poljak, Anne
114fe1b4-3275-4d81-8d9c-ff523cd18254
Guilhaus, Michael
f2e34a05-c65c-41cc-9f30-268d41d5d3e7
Cavicchioli, Ricardo
95c04c14-6cdd-4f0d-afc6-32bdbfa53ad9

Williams, Timothy J., Burg, Dominic W., Ertan, Haluk, Raftery, Mark J., Poljak, Anne, Guilhaus, Michael and Cavicchioli, Ricardo (2010) Global proteomic analysis of the insoluble, soluble, and supernatant fractions of the psychrophilic archaeon Methanococcoides burtonii. Part II: the effect of different methylated growth substrates. Journal of Proteome Research, 9 (2), 653-663. (doi:10.1021/pr9005102). (PMID:19947665)

Record type: Article

Abstract

Methanococcoides burtonii is a cold-adapted methanogenic archaeon from Ace Lake in Antarctica. Methanol and methylamines are the only substrates it can use for carbon and energy. We carried out quantitative proteomics using iTRAQ of M. burtonii cells grown on different substrates (methanol in defined media or trimethylamine in complex media), using techniques that enriched for secreted and membrane proteins in addition to cytoplasmic proteins. By integrating proteomic data with the complete, manually annotated genome sequence of M. burtonii, we were able to gain new insight into methylotrophic metabolism and the effects of methanol on the cell. Metabolic processing of methanol and methylamines is initiated by methyltransferases specific for each substrate, with multiple paralogs for each of the methyltransferases (similar to other members of the Methanosarcinaceae). In M. burtonii, most methyltransferases appear to have distinct roles in the metabolism of methylated substrates, although two methylamine methyltransferases appear to be nonfunctional. One set of methyltransferases for trimethylamine catabolism appears to be membrane associated, potentially providing a mechanism to directly couple trimethylamine uptake to demethylation. Important roles were highlighted for citrate synthase, glutamine synthetase, acetyl-CoA decarbonylase/synthase, and pyruvate synthase in carbon and nitrogen metabolism during growth on methanol. M. burtonii had only a marginal response to the provision of exogenous amino acids (from yeast extract), indicating that it is predisposed to the endogenous synthesis of amino acids. Growth on methanol appeared to cause oxidative stress in the cell, possibly through the formation of reactive nonoxygen species and formaldehyde, and the oxidative inactivation of corrinoid proteins, with the cell responding by elevating the synthesis of universal stress (Usp) proteins, several nucleic acid binding proteins, and a serpin. In addition, changes in levels of cell envelope proteins were linked to counteracting the disruptive solvent effects of methanol on cell membranes. This is the first global proteomic study to examine the effects of different carbon sources on the growth of an obligately methylotrophic methanogen.

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e-pub ahead of print date: 1 December 2009
Published date: 5 February 2010
Organisations: Molecular and Cellular

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Local EPrints ID: 338911
URI: http://eprints.soton.ac.uk/id/eprint/338911
ISSN: 1535-3893
PURE UUID: 8354ec3e-29e9-4002-9c6e-ac90e1c6fc4a

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Date deposited: 18 May 2012 13:20
Last modified: 14 Mar 2024 11:06

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Contributors

Author: Timothy J. Williams
Author: Dominic W. Burg
Author: Haluk Ertan
Author: Mark J. Raftery
Author: Anne Poljak
Author: Michael Guilhaus
Author: Ricardo Cavicchioli

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