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Methods Enzymol. 2011;494:257-80. doi: 10.1016/B978-0-12-385112-3.00013-5.

Proton translocation in methanogens.

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Institute of Microbiology and Biotechnology, University of Bonn, Bonn, Germany.


Methanogenic archaea of the genus Methanosarcina possess a unique type of metabolism because they use H(2)+CO(2), methylated C(1)-compounds, or acetate as energy and carbon source for growth. The process of methanogenesis is fundamental for the global carbon cycle and represents the terminal step in the anaerobic breakdown of organic matter in freshwater sediments. Moreover, methane is an important greenhouse gas that directly contributes to climate change and global warming. Methanosarcina species convert the aforementioned substrates to CH(4) via the CO(2)-reducing, the methylotrophic, or the aceticlastic pathway. All methanogenic processes finally result in the oxidation of two thiol-containing cofactors (HS-CoM and HS-CoB), leading to the formation of the so-called heterodisulfide (CoM-S-S-CoB) that contains an intermolecular disulfide bridge. This molecule functions as the terminal electron acceptor of a branched respiratory chain. Molecular hydrogen, reduced coenzyme F(420), or reduced ferredoxin are used as electron donors. The key enzymes of the respiratory chain (Ech hydrogenase, F(420)-nonreducing hydrogenase, F(420)H(2) dehydrogenase, and heterodisulfide reductase) couple the redox reactions to proton translocation across the cytoplasmic membrane. The resulting electrochemical proton gradient is the driving force for ATP synthesis. Here, we describe the methods and techniques of how to analyze electron transfer reactions, the process of proton translocation, and the formation of ATP.

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