X-ray Structure of the [FeFe]-Hydrogenase Maturase HydE from Thermotoga maritima

Nicolet, Yvain; Rubach, Jon K.; Posewitz, Matthew C.; Amara, Patricia; Mathevon, Carole; Atta, Mohamed; Fontecave, Marc; Fontecilla‐Camps, Juan C.

doi:10.1074/jbc.m801161200

Cited by 122 publications

(214 citation statements)

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“…3). This assumption is based on the X-ray structure of HydE that displays three anion-binding sites, one of which binds SCN À with good affinity [27].…”

Section: Resultsmentioning

confidence: 99%

“…They are also likely to belong to the subclass of Radical-SAM proteins characterized by a complete TIM-barrel and a buried active site suitable for small substrates [27]. Surprisingly, and contrary to the other members of the family, HydG and ThiH have similar patches of conserved amino acids mostly found at the internal faces of the b-strands that define the barrel ( Fig.…”

Section: Amino Acid Sequence Comparison Of Hydg and Thihmentioning

confidence: 99%

“…Peters et al have also hypothesized that the diatomic CO and CN ligands are made from glycine although this activity was not assigned to any maturase in particular [29]. Our own structural studies of HydE have shown that this protein has three, evenly spaced, anion-binding sites in a large cavity, inside the (ab) 8 barrel [27]. One of these sites binds SCN À with good affinity.…”

Section: Introductionmentioning

confidence: 99%

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The role of the maturase HydG in [FeFe]‐hydrogenase active site synthesis and assembly

et al. 2009

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a b s t r a c t[FeFe]-hydrogenases catalyze the protons/hydrogen interconversion through a unique di-iron active site consisting of three CO and two CN ligands, and a non-protein SCH 2 XCH 2 S (X = N or O) dithiolate bridge. Site assembly requires two ''Radical-S-adenosylmethionine (SAM or AdoMet)" iron-sulfur enzymes, HydE and HydG, and one GTPase, HydF. The sequence homology between HydG and ThiH, a Radical-SAM enzyme which cleaves tyrosine into p-cresol and dehydroglycine, and the finding of a similar cleavage reaction catalyzed by HydG suggests a mechanism for hydrogenase maturation. Here we propose that HydG is specifically involved in the synthesis of the dithiolate ligand, with two tyrosine-derived dehydroglycines as precursors along with an [FeS] cluster of HydG functioning both as electron shuttle and source of the sulfur atoms.

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“…3). This assumption is based on the X-ray structure of HydE that displays three anion-binding sites, one of which binds SCN À with good affinity [27].…”

Section: Resultsmentioning

confidence: 99%

Section: Amino Acid Sequence Comparison Of Hydg and Thihmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The role of the maturase HydG in [FeFe]‐hydrogenase active site synthesis and assembly

et al. 2009

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“…Several years ago, structures of TmHydE were reported from a crystal form that diffracted to high resolution (1.25 Å) [30]. These crystals have been used in a crystallographic study of the reductive cleavage of SAM [31].…”

Section: Hyde: 'A Riddle Wrapped In a Mystery Inside An Enigma'mentioning

confidence: 99%

“…These crystals have been used in a crystallographic study of the reductive cleavage of SAM [31]. Some HydE sequences contain an auxiliary cluster binding site, but it is of variable constitution and can be removed by mutation without inactivating HydE, suggesting it is functionally inessential [30,32]. The structure of HydE (Figure 4a) includes a large cavity (991 Å 3 ) within the TIM barrel that is well suited to bind a second substrate (in addition to SAM).…”

Section: Hyde: 'A Riddle Wrapped In a Mystery Inside An Enigma'mentioning

confidence: 99%

Metallocofactor assembly for [FeFe]-hydrogenases

Dinis

Wieckowski

Roach

2016

Current Opinion in Structural Biology

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Hydrogenases are a potential source of environmentally benign bioenergy, using complex cofactors to catalyze the reversible reduction of protons to form hydrogen. AddressesChemistry and the Institute for Life Sciences, University of Southampton, Highfield Campus, Southampton SO17 1BJ, UK.Corresponding author: Peter L. Roach (plr2@soton.ac.uk). AbstractHydrogenases are a potential source of environmentally benign bioenergy, using complex cofactors to catalyze the reversible reduction of protons to form hydrogen. The most active subclass, the [FeFe]-hydrogenases, is dependent on a metallocofactor, the H cluster, that consists of a two iron subcluster ([2Fe] H ) bridging to a classical cubane cluster ([4Fe-4S] H ). The ligands coordinating to the diiron subcluster include an azadithiolate, three carbon monoxides, and two cyanides. To assemble this complex cofactor, three maturase enzymes, HydG, HydE and HydF are required. The biosynthesis of the diatomic ligands proceeds by an unusual fragmentation mechanism, and structural studies in combination with spectroscopic analysis have started to provide insights into the HydG mediated assembly of a [2Fe] H subcluster precursor.

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[ FeFe ]‐Hydrogenase Cofactor Assembly

Shepard

Byer

Boyd

et al. 2004

Encyclopedia of Inorganic and Bioinorganic Chemistry

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This article highlights recent advances in our understanding of the biosynthetic pathway for active site H‐cluster assembly in [ FeFe ]‐hydrogenases. The H‐cluster is composed of a [ 4Fe–4S ] cubane bridged via one cysteine thiolate to a 2Fe subcluster; the 2Fe subcluster is further coordinated by carbon monoxide, cyanide, and bridging dithiolate ligands. Biosynthesis of the H‐cluster occurs through stepwise modification of simple Fe–S cluster precursors; these chemical modifications are carried out by three gene products denoted HydE , HydF , and HydG . Maturation of the [ FeFe ]‐hydrogenase requires the presence of a preformed [ 4Fe–4S ] cluster, indicating that HydE , HydF , and HydG are directed toward the biosynthesis of the 2Fe subcluster. HydE and HydG are both radical S ‐adenosylmethionine ( SAM ) enzymes and utilize a CX 3 CX 2 C motif to bind site‐differentiated [ 4Fe–4S ] 2+/+ clusters that promote the reductive cleavage of SAM into methionine and a 5′‐deoxyadenosyl radical responsible for hydrogen atom abstraction from substrate. In an extraordinary biochemical reaction, HydG uses this chemistry to catalyze the radical degradation of tyrosine into p ‐cresol and a glycine‐like intermediate; an accessory, C‐terminal [ 4Fe–4S ] cluster then degrades the latter intermediate into cyanide and carbon monoxide. HydE is presumed to be responsible for bridging dithiolate ligand biosynthesis from an unknown substrate. HydF contains an N‐terminal GTPase domain and a C‐terminal Fe–S ‐cluster‐binding domain and binds both [ 2Fe–2S ] and [ 4Fe–4S ] clusters; evidence suggests that the former cluster is modified by HydE and HydG to form a 2Fe H‐cluster precursor on HydF . The role of GTP hydrolysis in H‐cluster maturation remains largely unexplained; however, it may be related to gating the protein–protein interactions between HydF and HydE and HydG . The reactions catalyzed by HydE and HydG can be thought of as specific ligand modification events designed to fine‐tune H‐cluster reactivity; not only do the chemical transformations themselves have direct links to reactions important on early Earth, but radical SAM enzymes with their canonical CX 3 CX 2 C motif are likely very ancient. The modern diversity that is observed in chemical reactions catalyzed by the radical SAM superfamily can be explained evolutionarily through the recruitment of distinct protein domains that impart specific substrate activation capabilities. Recruitment of these domains is discussed in the context of HydE and HydG and important links are made to the nitrogenase system where intriguing parallels between [ FeFe ]‐hydA H‐cluster biosynthesis and nitrogenase FeMo ‐cofactor assembly clearly exist.

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X-ray Structure of the [FeFe]-Hydrogenase Maturase HydE from Thermotoga maritima

Abstract: ؊ with high affinity. Although the overall triose-phosphate isomerase-barrel structure of HydE is very similar to that of biotin synthase, the residues that line the internal cavity are significantly different in the two enzymes.

Cited by 122 publications

References 57 publications

The role of the maturase HydG in [FeFe]‐hydrogenase active site synthesis and assembly

The role of the maturase HydG in [FeFe]‐hydrogenase active site synthesis and assembly

Metallocofactor assembly for [FeFe]-hydrogenases

[ FeFe ]‐Hydrogenase Cofactor Assembly

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