Isolation of an enzyme complex with carbon monoxide dehydrogenase activity containing corrinoid and nickel from acetate-grown Methanosarcina thermophila

Terlesky, Katherine C.; Nelson, Michael J.; Ferry, James G.

doi:10.1128/jb.168.3.1053-1058.1986

Cited by 154 publications

(116 citation statements)

References 30 publications

Supporting

Mentioning

110

Contrasting

Order By: Relevance

“…Although the CooS paralog in R. rubrum reduces ferredoxin (33), the enzyme in M. acetivorans has not been investigated, and the electron acceptor is unknown. The genome also is annotated (www.tigr.org) with duplicate gene clusters (MA1011-MA1016 and MA3860-MA3865), each encoding paralogs of a five-subunit CO dehydrogenase͞ acetyl-CoA synthase (CdhABCDE) complex first described in Methanosarcina thermophila (34,35). Three subunits (MA1014-MA1016 and MA3860-MA3862) from each complex were detected and found to be elevated an average of 20-fold in CO-grown versus methanol-grown cells in contrast to an Ϸ2-fold differential abundance versus acetate-grown cells ( Table 1).…”

Section: Resultsmentioning

confidence: 99%

An unconventional pathway for reduction of CO ₂ to methane in CO-grown Methanosarcina acetivorans revealed by proteomics

Lessner

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

120

154

View full text Add to dashboard Cite

The results indicate that oxidation of CO to CO 2 supplies electrons for reduction of CO 2 to a methyl group by steps and enzymes of the pathway for CO 2 reduction determined for other methane-producing species. However, proteomic and quantitative RT-PCR results suggest that reduction of the methyl group to methane involves novel methyltransferases and a coenzyme F 420H2:heterodisulfide oxidoreductase system that generates a proton gradient for ATP synthesis not previously described for pathways reducing CO 2 to methane. Biochemical assays support a role for the oxidoreductase, and transcriptional mapping identified an unusual operon structure encoding the oxidoreductase. The proteomic results further indicate that acetate is synthesized from the methyl group and CO by a reversal of initial steps in the pathway for conversion of acetate to methane that yields ATP by substrate level phosphorylation. The results indicate that M. acetivorans utilizes a pathway distinct from all known CO 2 reduction pathways for methane formation that reflects an adaptation to the marine environment. Finally, the pathway supports the basis for a recently proposed primitive CO-dependent energy-conservation cycle that drove and directed the early evolution of life on Earth.anaerobic ͉ Archaea ͉ carbon monoxide C arbon monoxide (CO), an atmospheric pollutant that binds tightly to hemoglobin, is held below toxic levels in part by both aerobic and anaerobic microbes (1). The microbial metabolism of CO is an important component of the global carbon cycle (1, 2), and CO is believed to have been present in the atmosphere of early Earth that fueled the evolution of primitive metabolisms (3-7). Investigations of aerobic species from the Bacteria domain have contributed important insights into microbial CO oxidation (8, 9), as have investigations of anaerobes from the Bacteria domain that conserve energy by coupling CO oxidation to H 2 evolution (10-12). Further understanding has been derived from studies of CO-using anaerobes from the Bacteria domain that conserve energy by oxidizing CO and reducing CO 2 to acetate (13,14) or reducing sulfate to sulfide (15). Far less is known for pathways of the few CO-using species in the Archaea domain that have been described. Methanothermobacter thermautotrophicus, Methanosarcina barkeri, and Methanosarcina acetivorans obtain energy for growth by converting CO to methane (16)(17)(18)(19)(20). Although methane formation from CO first was reported in 1947 (21), a comprehensive understanding of the overall pathway for any species has not been reported. It is postulated that M. barkeri oxidizes CO to H 2 , and the H 2 is reoxidized to provide electrons for reducing CO 2 to methane (16). It is postulated further that H 2 production is essential for ATP synthesis during growth on CO (16,22,23). M. acetivorans was isolated from marine sediments where giant kelp is decomposed to methane (24). The flotation bladders of kelp contain CO that is a presumed substrate for M. acetivorans in nature. M. acetivorans produ...

show abstract

Section: Resultsmentioning

confidence: 99%

An unconventional pathway for reduction of CO ₂ to methane in CO-grown Methanosarcina acetivorans revealed by proteomics

Lessner

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

120

154

View full text Add to dashboard Cite

show abstract

“…Glycerol was added to a final concentration of 10% (wt/vol) before the enzyme was stored in liquid N2. The carbon monoxide dehydrogenase (CODH) enzyme complex (41), the nickel/iron-sulfur and corrinoid/iron-sulfur components of the complex (2), and ferredoxin (39) were purified as described previously.…”

Section: Methodsmentioning

confidence: 99%

Purification and properties of methyl coenzyme M methylreductase from acetate-grown Methanosarcina thermophila

Jablonski

Ferry

1991

J Bacteriol

Self Cite

View full text Add to dashboard Cite

Methyl coenzyme M methylreductase from acetate-grown Methanosarcina thermophila TM-1 was purified 16-fold from a cell extract to apparent homogeneity as determined by native polyacrylamide gel electrophoresis. Ninety-four percent of the methylreductase activity was recovered in the soluble fraction of cell extracts. The estimated native molecular weight of the enzyme was between 132,000 (standard deviation [SD], 1,200) and 141,000 (SD,1,200). Denaturing polyacrylamide gel electrophoresis revealed three protein bands corresponding to molecular weights of 69,000 (SD,1,200),42,000 (SD,1,200), and 33,000 (SD,1,200) and indicated a subunit configuration of alp'lyl. As isolated, the enzyme was inactive but could be reductively reactivated with titanium (III) citrate or reduced ferredoxin. ATP stimulated enzyme reactivation and was postulated to be involved in a conformational change of the inactive enzyme from an unready state to a ready state that could be reductively reactivated. The temperature and pH optima for enzyme activity were 60°C and between 6.5 and 7.0, respectively. The active enzyme contained 1 mol of coenzyme F430 per mol of enzyme (M,r 144,000). The Kms for 2-(methylthio)ethane-sulfonate and 7-mercaptoheptanoylthreonine phosphate were 3.3 mM and 59,uM, respectively.Methanogenic bacteria obtain energy for growth by reducing carbon dioxide to methane or converting the methyl group of acetate, methanol, or methylated amines to methane. 2-(Methylthio)ethane-sulfonate (CH3-S-CoM) is a common intermediate that is reductively demethylated to CH4 by methyl coenzyme M methylreductase. The extensively studied methylreductases have a native molecular weight of approximately 300,000 and a subunit composition of a212Y2.In addition, these methylreductases contain 2 mol of a nickel-containing tetrahydrocorphin (coenzyme F430) per mol of enzyme (Mr, 300,000). The methylreductase from Methanobacterium thermoautotrophicum AH is the most extensively characterized (11, 14, 15, 26, 29-31, 33, 34, 36); the enzyme is one component of the H2-dependent methylreductase system, which has been reconstituted with protein fractions Al, A3a, and A3b, purified protein A2, and C (the methylreductase) (26,34,36). Only components A2 (34) and the methylreductase (15) have been purified to homogeneity. As isolated, the methylreductase must first be reactivated which is thought to occur by reduction of Ni(II) to Ni(I) in F430 (4). Reductive reactivation requires components A3a, A2, and ATP. H2 is oxidized by a hydrogenase in component A3b to provide electrons for the reactivation. Titanium (III) citrate can replace H2 and component A3b as an alternative source of electrons. Once reactivated, the electron donor to the methylreductase is 7-mercaptoheptanoylthreonine phosphate (HS-HTP) and the products of the reaction are CH4 and the heterodisulfide of HS-CoM and HS-HTP. The heterodisulfide is then reduced by H2 and component Al to 2-mercaptoethanesulfonate (HS-CoM) and HS-HTP.Methyl coenzyme M methylreductases from Methanosarcina spp. have ...

show abstract

“…The central enzyme in this pathway is acetyl-CoA decarbonylase/synthase (ACDS), a multimeric 2.4-MDa ␣ 8 ␤ 8 ␥ 8 ␦ 8 8 complex that mediates multiple reactions (2,3). In Methanosarcina barkeri, the ␣ to subunits have sizes of 89, 60, 50, 48, and 20 kDa, respectively (4,5).…”

mentioning

confidence: 99%

Structure of the α ₂ ε ₂ Ni-dependent CO dehydrogenase component of the Methanosarcina barkeri acetyl-CoA decarbonylase/synthase complex

Gong

Hao

Wei

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

114

119

View full text Add to dashboard Cite

Ni-dependent carbon monoxide dehydrogenases (Ni-CODHs) are a diverse family of enzymes that catalyze reversible CO:CO2 oxidoreductase activity in acetogens, methanogens, and some COusing bacteria. Crystallography of Ni-CODHs from CO-using bacteria and acetogens has revealed the overall fold of the Ni-CODH core and has suggested structures for the C cluster that mediates CO:CO2 interconversion. Despite these advances, the mechanism of CO oxidation has remained elusive. Herein, we report the structure of a distinct class of Ni-CODH from methanogenic archaea: the ␣2 2 component from the ␣8␤8␥8␦8 8 CODH/acetyl-CoA decarbonylase/ synthase complex, an enzyme responsible for the majority of biogenic methane production on Earth. The structure of this Ni-CODH component provides support for a hitherto unobserved state in which both CO and H2O/OH -bind to the Ni and the exogenous FCII iron of the C cluster, respectively, and offers insight into the structures and functional roles of the -subunit and FeS domain not present in nonmethanogenic Ni-CODHs.carbon dioxide ͉ carbon monoxide ͉ oxidoreductase ͉ acetate ͉ methanogenesis

show abstract

Isolation of an enzyme complex with carbon monoxide dehydrogenase activity containing corrinoid and nickel from acetate-grown Methanosarcina thermophila

Cited by 154 publications

References 30 publications

An unconventional pathway for reduction of CO ₂ to methane in CO-grown Methanosarcina acetivorans revealed by proteomics

An unconventional pathway for reduction of CO ₂ to methane in CO-grown Methanosarcina acetivorans revealed by proteomics

Purification and properties of methyl coenzyme M methylreductase from acetate-grown Methanosarcina thermophila

Structure of the α ₂ ε ₂ Ni-dependent CO dehydrogenase component of the Methanosarcina barkeri acetyl-CoA decarbonylase/synthase complex

Contact Info

Product

Resources

About

Isolation of an enzyme complex with carbon monoxide dehydrogenase activity containing corrinoid and nickel from acetate-grown Methanosarcina thermophila

Cited by 154 publications

References 30 publications

An unconventional pathway for reduction of CO 2 to methane in CO-grown Methanosarcina acetivorans revealed by proteomics

An unconventional pathway for reduction of CO 2 to methane in CO-grown Methanosarcina acetivorans revealed by proteomics

Purification and properties of methyl coenzyme M methylreductase from acetate-grown Methanosarcina thermophila

Structure of the α 2 ε 2 Ni-dependent CO dehydrogenase component of the Methanosarcina barkeri acetyl-CoA decarbonylase/synthase complex

Contact Info

Product

Resources

About

An unconventional pathway for reduction of CO ₂ to methane in CO-grown Methanosarcina acetivorans revealed by proteomics

An unconventional pathway for reduction of CO ₂ to methane in CO-grown Methanosarcina acetivorans revealed by proteomics

Structure of the α ₂ ε ₂ Ni-dependent CO dehydrogenase component of the Methanosarcina barkeri acetyl-CoA decarbonylase/synthase complex