A1Ao-ATP Synthase of Methanobrevibacter ruminantium Couples Sodium Ions for ATP Synthesis under Physiological Conditions

McMillan, Duncan G. G.; Ferguson, Scott; Dey, Debjit; Schröder, Katja; Aung, Htin; Carbone, Vincenzo; Attwood, Graeme T.; Ronimus, Ron S.; Meier, Thomas; Janssen, Peter H.; Cook, Gregory M.

doi:10.1074/jbc.m111.281675

Cited by 38 publications

(36 citation statements)

References 69 publications

(69 reference statements)

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“…strain AbM4 (Table 2). Monensin, a sodium ionophore, and 2-bromoethanesulfonic acid (BES), an analogue of methyl-coenzyme M, are potent inhibitors of methanogens (34). Using monensin (1 M) or BES (30 M), nearly complete inhibition, i.e., Ͼ85% reduction of growth, was observed for both strains (Tables 1 and 2).…”

Section: Resultsmentioning

confidence: 92%

“…Methanococcus maripaludis strain S2 is a well-characterized genetically tractable methanogen that can be grown in the absence of H 2 and CO 2 using formate (27)(28)(29)(30)(31)(32). M. maripaludis grows quickly to high cell densities in contrast to slowly growing rumen isolates, such as Methanobrevibacter ruminantium M1, where cell densities are low (33,34). Both methanogens are typically grown using anaerobic culturing techniques in 5-ml or greater culture volumes using appropriately sealed and pressurized glass tubes, which is incompatible with modern high-throughput screening techniques for drug discovery and phenotypic analysis (35).…”

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confidence: 99%

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Development of Multiwell-Plate Methods Using Pure Cultures of Methanogens To Identify New Inhibitors for Suppressing Ruminant Methane Emissions

Weimar

Cheung

Dey

et al. 2017

Appl Environ Microbiol

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Hydrogenotrophic methanogens typically require strictly anaerobic culturing conditions in glass tubes with overpressures of H 2 and CO 2 that are both time-consuming and costly. To increase the throughput for screening chemical compound libraries, 96-well microtiter plate methods for the growth of a marine (environmental) methanogen Methanococcus maripaludis strain S2 and the rumen methanogen Methanobrevibacter species AbM4 were developed. A number of key parameters (inoculum size, reducing agents for medium preparation, assay duration, inhibitor solvents, and culture volume) were optimized to achieve robust and reproducible growth in a high-throughput microtiter plate format. The method was validated using published methanogen inhibitors and statistically assessed for sensitivity and reproducibility. The Sigma-Aldrich LOPAC library containing 1,280 pharmacologically active compounds and an in-house natural product library (120 compounds) were screened against M. maripaludis as a proof of utility. This screen identified a number of bioactive compounds, and MIC values were confirmed for some of them against M. maripaludis and M. AbM4. The developed method provides a significant increase in throughput for screening compound libraries and can now be used to screen larger compound libraries to discover novel methanogen-specific inhibitors for the mitigation of ruminant methane emissions.IMPORTANCE Methane emissions from ruminants are a significant contributor to global greenhouse gas emissions, and new technologies are required to control emissions in the agriculture technology (agritech) sector. The discovery of small-molecule inhibitors of methanogens using high-throughput phenotypic (growth) screening against compound libraries (synthetic and natural products) is an attractive avenue. However, phenotypic inhibitor screening is currently hindered by our inability to grow methanogens in a high-throughput format. We have developed, optimized, and validated a highthroughput 96-well microtiter plate assay for growing environmental and rumen methanogens. Using this platform, we identified several new inhibitors of methanogen growth, demonstrating the utility of this approach to fast track the development of methanogen-specific inhibitors for controlling ruminant methane emissions.KEYWORDS methanogen, greenhouse gas, Methanococcus maripaludis, highthroughput, rumen, Methanobrevibacter M ethane emissions from ruminants are a significant contributor to global greenhouse gas emissions (1). In countries such as New Zealand, with a large pasturebased livestock sector, greenhouse gas emissions from agriculture represent approxi-

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Section: Resultsmentioning

confidence: 92%

mentioning

confidence: 99%

Development of Multiwell-Plate Methods Using Pure Cultures of Methanogens To Identify New Inhibitors for Suppressing Ruminant Methane Emissions

Weimar

Cheung

Dey

et al. 2017

Appl Environ Microbiol

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“…Further, the A 1 A 0 ATPase/synthases studied among the members of the groups Methanobacteriales/Methanococcales have clearly been shown to pump Na + (McMillan et al, 2011;Mulkidjanian et al, 2008). In contrast, the cytochrome-containing methanogens in the Methanosarcinales have Mtr complexes, but also have steps that pump protons (Schlegel & Müller, 2013;Thauer et al, 2008), and evidence has favoured H + pumping by the ATPases in these organisms (Müller et al, 1999;Pisa et al, 2007).…”

Section: Adaptation To High Proton Concentrationsmentioning

confidence: 99%

“…In Table 3, partial sequences of the AtpC/K are aligned, and residues critical to Na + binding in the Methanobacteriales/Methanococcales (Grüber et al, 2014;McMillan et al, 2011) are indicated in bold. Also shown in bold are residues in the M. acetivorans sequence that are considered crucial to being able to bind either Na + or H + .…”

Section: Adaptation To High Proton Concentrationsmentioning

confidence: 99%

Genome of Methanoregula boonei 6A8 reveals adaptations to oligotrophic peatland environments

et al. 2015

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Analysis of the genome sequence of Methanoregula boonei strain 6A8, an acidophilic methanogen isolated from an ombrotrophic (rain-fed) peat bog, has revealed unique features that likely allow it to survive in acidic, nutrient-poor conditions. First, M. boonei is predicted to generate ATP using protons that are abundant in peat, rather than sodium ions that are scarce, and the sequence of a membrane-bound methyltransferase, believed to pump Na + in all methanogens, shows differences in key amino acid residues. Further, perhaps reflecting the hypokalemic status of many peat bogs, M. boonei demonstrates redundancy in the predicted potassium uptake genes trk, kdp and kup, some of which may have been horizontally transferred to methanogens from bacteria, possibly Geobacter spp. Overall, the putative functions of the potassium uptake, ATPase and methyltransferase genes may, at least in part, explain the cosmopolitan success of group E1/E2 and related methanogenic archaea in acidic peat bogs.

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“…A switch from Na 1 to H 1 pumping has been observed for the ATP synthases from Propionigenium modestum and Methanobrevibacter ruminantium at low pH and low levels of Na 1 (47,48). This can be explained by the weak H 1 -selectivity of these ATP synthases as demonstrated by Krah and coworkers (49).…”

Section: The Possibility Of Na 1 Being a Coupling Ion In Complex Imentioning

confidence: 81%

The role of proton and sodium ions in energy transduction by respiratory complex I

2012

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SummaryRespiratory complex I plays a central role in energy transduction. It catalyzes the oxidation of NADH and the reduction of quinone, coupled to cation translocation across the membrane, thereby establishing an electrochemical potential. For more than half a century, data on complex I has been gathered, including recently determined crystal structures, yet complex I is the least understood complex of the respiratory chain. The mechanisms of quinone reduction, charge translocation and their coupling are still unknown. The H 1 is accepted to be the coupling ion of the system; however, Na 1 has also been suggested to perform such a role. In this article, we address the relation of those two ions with complex I and refer ion pump and Na 1 /H 1 antiporter as possible transport mechanisms of the system. We put forward a hypothesis to explain some apparently contradictory data on the nature of the coupling ion, and we revisit the role of H 1 and Na 1 cycles in the overall bioenergetics of the cell. IUBMBIUBMB Life, 64(6): 492-498, 2012

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A1Ao-ATP Synthase of Methanobrevibacter ruminantium Couples Sodium Ions for ATP Synthesis under Physiological Conditions

Cited by 38 publications

References 69 publications

Development of Multiwell-Plate Methods Using Pure Cultures of Methanogens To Identify New Inhibitors for Suppressing Ruminant Methane Emissions

Development of Multiwell-Plate Methods Using Pure Cultures of Methanogens To Identify New Inhibitors for Suppressing Ruminant Methane Emissions

Genome of Methanoregula boonei 6A8 reveals adaptations to oligotrophic peatland environments

The role of proton and sodium ions in energy transduction by respiratory complex I

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