Reaction Mechanism and Structural Model of ADP-forming Acetyl-CoA Synthetase from the Hyperthermophilic Archaeon Pyrococcus furiosus

Bräsen, Christopher; Schmidt, Marcel; Grötzinger, Joachim; Schönheit, Peter

doi:10.1074/jbc.m710218200

Cited by 38 publications

(52 citation statements)

References 38 publications

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“…The predicted beta-subunit of at least one of the putative acetyl-CoA synthetases contained a conserved histidine residue typical of well characterised acetyl-CoA synthetases that specifically form acetate and differentiate them from other structurally related succinyl-CoA synthetases (Bräsen et al, 2008). DEH-J10 may therefore gain ATP by substrate-level phosphorylation during the conversion of acetyl-CoA to acetate (McInerney et al, 2007;Bräsen et al, 2008).…”

Section: Acetyl-coa Synthetasesmentioning

confidence: 99%

“…Most of the top BLASTP hits for these predicted proteins were to proteins from the archaeal genera Pyrococcus and Thermococcus, which are known to catalyse the one-step formation of acetate from acetyl-CoA with the concomitant phosphorylation of ADP to ATP (Bräsen et al, 2008). The predicted beta-subunit of at least one of the putative acetyl-CoA synthetases contained a conserved histidine residue typical of well characterised acetyl-CoA synthetases that specifically form acetate and differentiate them from other structurally related succinyl-CoA synthetases (Bräsen et al, 2008). DEH-J10 may therefore gain ATP by substrate-level phosphorylation during the conversion of acetyl-CoA to acetate (McInerney et al, 2007;Bräsen et al, 2008).…”

Section: Acetyl-coa Synthetasesmentioning

confidence: 99%

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Genome sequencing of a single cell of the widely distributed marine subsurface Dehalococcoidia, phylum Chloroflexi

et al. 2013

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Bacteria of the class Dehalococcoidia (DEH), phylum Chloroflexi, are widely distributed in the marine subsurface, yet metabolic properties of the many uncultivated lineages are completely unknown. This study therefore analysed genomic content from a single DEH cell designated 'DEH-J10' obtained from the sediments of Aarhus Bay, Denmark. Real-time PCR showed the DEH-J10 phylotype was abundant in upper sediments but was absent below 160 cm below sea floor. A 1.44 Mbp assembly was obtained and was estimated to represent up to 60.8% of the full genome. The predicted genome is much larger than genomes of cultivated DEH and appears to confer metabolic versatility. Numerous genes encoding enzymes of core and auxiliary beta-oxidation pathways were identified, suggesting that this organism is capable of oxidising various fatty acids and/or structurally related substrates. Additional substrate versatility was indicated by genes, which may enable the bacterium to oxidise aromatic compounds. Genes encoding enzymes of the reductive acetyl-CoA pathway were identified, which may also enable the fixation of CO 2 or oxidation of organics completely to CO 2 . Genes encoding a putative dimethylsulphoxide reductase were the only evidence for a respiratory terminal reductase. No evidence for reductive dehalogenase genes was found. Genetic evidence also suggests that the organism could synthesise ATP by converting acetyl-CoA to acetate by substrate-level phosphorylation. Other encoded enzymes putatively conferring marine adaptations such as salt tolerance and organo-sulphate sulfohydrolysis were identified. Together, these analyses provide the first insights into the potential metabolic traits that may enable members of the DEH to occupy an ecological niche in marine sediments.

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Section: Acetyl-coa Synthetasesmentioning

confidence: 99%

Section: Acetyl-coa Synthetasesmentioning

confidence: 99%

Genome sequencing of a single cell of the widely distributed marine subsurface Dehalococcoidia, phylum Chloroflexi

et al. 2013

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“…A mechanism comparable to the mechanism of the SCSs was presumed for the ACDs (20). A study based on sequence, biochemical, and mutational analyses identified a highly conserved and functional relevant His residue within the beta subunit of the ACDs (20). Of note, the SCS enzymes do not contain a comparable His residue.…”

mentioning

confidence: 99%

“…Of note, the SCS enzymes do not contain a comparable His residue. Thus, an extension of the mechanism by a fourth step was suggested for the ACD1 from Pyrococcus furiosus (20). Here, the second His residue is thought to serve as an additional intermediate, which gets phosphorylated transiently during the enzymatic reaction and subsequently transmits the phosphoryl moiety onto the bound NDP.…”

mentioning

confidence: 99%

“…The distance of around 36 Å between site I and site II is assumed to be bridged by a so-called "swinging loop" (12,14,19). A mechanism comparable to the mechanism of the SCSs was presumed for the ACDs (20). A study based on sequence, biochemical, and mutational analyses identified a highly conserved and functional relevant His residue within the beta subunit of the ACDs (20).…”

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confidence: 99%

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Structure of NDP-forming Acetyl-CoA synthetase ACD1 reveals a large rearrangement for phosphoryl transfer

Weisse

Faust

Schmidt

et al. 2016

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

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The NDP-forming acyl-CoA synthetases (ACDs) catalyze the conversion of various CoA thioesters to the corresponding acids, conserving their chemical energy in form of ATP. The ACDs are the major energy-conserving enzymes in sugar and peptide fermentation of hyperthermophilic archaea. They are considered to be primordial enzymes of ATP synthesis in the early evolution of life. We present the first crystal structures, to our knowledge, of an ACD from the hyperthermophilic archaeon Candidatus Korachaeum cryptofilum. These structures reveal a unique arrangement of the ACD subunits alpha and beta within an α 2 β 2 -heterotetrameric complex. This arrangement significantly differs from other members of the superfamily. To transmit an activated phosphoryl moiety from the Ac-CoA binding site (within the alpha subunit) to the NDP-binding site (within the beta subunit), a distance of 51 Å has to be bridged. This transmission requires a larger rearrangement within the protein complex involving a 21-aa-long phosphohistidinecontaining segment of the alpha subunit. Spatial restraints of the interaction of this segment with the beta subunit explain the necessity for a second highly conserved His residue within the beta subunit. The data support the proposed four-step reaction mechanism of ACDs, coupling acyl-CoA thioesters with ATP synthesis. Furthermore, the determined crystal structure of the complex with bound Ac-CoA allows first insight, to our knowledge, into the determinants for acyl-CoA substrate specificity. The composition and size of loops protruding into the binding pocket of acyl-CoA are determined by the individual arrangement of the characteristic subdomains.X-ray structure | metabolic energy conversion | protein dynamics | acyl-coenzyme A thioester | superfamily N DP-forming acyl-CoA synthetases (ACDs) catalyze the conversion of acyl-CoA thioesters to the corresponding acids and couple this reaction with the synthesis of ATP via the mechanism of substrate-level phosphorylation. ACDs have been studied in detail in hyperthermophilic archaea, where they function as the major energy-conserving enzymes in the course of anaerobic sugar and peptide fermentation (1-4). It is believed that ACDs represent a primordial mechanism of ATP synthesis in the early evolution of life. ACDs were found in all acetate (acid)-forming archaea (5, 6) and in the eukaryotic parasitic protists Entamoeba histolytica (7) and Giardia lamblia (8), but they have not been found in acetate-forming bacteria. In bacteria, with the exception of Chloroflexus (9), the conversion of inorganic phosphate and the thioester acetyl (Ac)-CoA to acetate and ATP is catalyzed by two enzymes, phosphate Ac-transferase and acetate kinase (10).

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Investigating the mechanism of ADP‐forming acetyl‐CoA synthetase from the protozoan parasite Entamoeba histolytica

2017

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ADP-forming acetyl-CoA synthetase (ACD) catalyzes the interconversion of acetyl-CoA and acetate. The related succinyl-CoA synthetase follows a three-step mechanism involving a single phosphoenzyme, but a novel four-step mechanism with two phosphoenzyme intermediates was proposed for Pyrococcus ACD. Characterization of enzyme variants of Entamoeba ACD in which the two proposed phosphorylated His residues were individually altered revealed that only His252 is essential for enzymatic activity. Analysis of variants altered at two residues proposed to interact with the phosphohistidine loop that swings between distinct parts of the active site are consistent with a mechanism involving a single phosphoenzyme intermediate. Our results suggest ACDs with different subunit structures may employ slightly different mechanisms to bridge the span between active sites I and II.

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Reaction Mechanism and Structural Model of ADP-forming Acetyl-CoA Synthetase from the Hyperthermophilic Archaeon Pyrococcus furiosus

Cited by 38 publications

References 38 publications

Genome sequencing of a single cell of the widely distributed marine subsurface Dehalococcoidia, phylum Chloroflexi

Genome sequencing of a single cell of the widely distributed marine subsurface Dehalococcoidia, phylum Chloroflexi

Structure of NDP-forming Acetyl-CoA synthetase ACD1 reveals a large rearrangement for phosphoryl transfer

Investigating the mechanism of ADP‐forming acetyl‐CoA synthetase from the protozoan parasite Entamoeba histolytica

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