Biochemical Analysis of the Interactions between the Proteins Involved in the [FeFe]-Hydrogenase Maturation Process

Vallese, Francesca; Berto, Paola; Ruzzene, Maria; Cendron, Laura; Sarno, Stefania; Rosa, Edith De; Giacometti, Giorgio M.; Costantini, Paola

doi:10.1074/jbc.m112.388900

Cited by 36 publications

(47 citation statements)

References 25 publications

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“…These results corroborate recent data showing that Clostridium acetobutylicum [FeFe] hydrogenase can interact with HydF only when co-expressed with the other two maturation factors (40).…”

Section: Hydrogenase Maturation Factor Interaction Profilessupporting

confidence: 82%

“…In total, our yeast two-hybrid library screens resulted in the identification of 1204 prey clones capable of supporting yeast growth when co-expressed with at least one of the FDX isoforms (supplemental Tables S2-S7; FDX1, 40;and FDX6,11). Of the putative interaction partners, 79 proteins were identified as potential interaction partners with more than one FDX isoform ( Table 1).…”

Section: Yeast Two-hybrid Library Screenmentioning

confidence: 99%

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Identification of Global Ferredoxin Interaction Networks in Chlamydomonas reinhardtii

Peden

Boehm

Mulder

et al. 2013

Journal of Biological Chemistry

100

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Background: Chlamydomonas contains six chloroplast ferredoxins (FDXs) whose function is still unclear. Results: A global FDX interactome was obtained where FDX1 has a predominant role and is the most relevant electron donor to FNR1 and HYDA1. Conclusion: FDXs have distinct but also overlapping function. Significance: We discovered new FDX interaction partners and specific roles for each FDX isoform.

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“…These results corroborate recent data showing that Clostridium acetobutylicum [FeFe] hydrogenase can interact with HydF only when co-expressed with the other two maturation factors (40).…”

Section: Hydrogenase Maturation Factor Interaction Profilessupporting

confidence: 82%

Section: Yeast Two-hybrid Library Screenmentioning

confidence: 99%

Identification of Global Ferredoxin Interaction Networks in Chlamydomonas reinhardtii

Peden

Boehm

Mulder

et al. 2013

Journal of Biological Chemistry

100

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“…Both HydG and HydE have been observed to stimulate the rate of GTP hydrolysis in HydF (38), possibly implying a connection between GTP hydrolysis on HydF and the interactions with the other two maturases. Surface plasmon resonance experiments corroborated this result, as the presence of GTP increased the rate of dissociation of HydG and HydE from HydF (65). Furthermore, although surface plasmon resonance experiments revealed that neither HydG nor HydE could displace each other from HydF, a stronger interaction between HydE and HydF compared with that between HydG and HydF was observed (65).…”

Section: Figure 4 Schematic Illustration Of the Reactions Catalyzed supporting

confidence: 61%

Radical S-Adenosyl-l-methionine Chemistry in the Synthesis of Hydrogenase and Nitrogenase Metal Cofactors

Byer

Shepard

Peters

et al. 2015

Journal of Biological Chemistry

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Despite the potential deleterious effects of radical reactions and unconstrained metal ions, the exquisitely controlled protein armature of metalloenzymes and the orchestration of their biosynthetic accessory proteins allow for the creation of complex metal centers that accomplish formidable catalysis. N 2 reduction to ammonia occurs at the nitrogenase FeMo cofactor, which can be viewed as a complex bridged metal assembly consisting of a [4Fe-3S] cluster linked to a [Mo-3Fe-3S] cluster by three sulfides and a central carbon (Figs. 1A and 2A) (1, 2). Reversible reduction of protons to H 2 occurs at the [FeFe]-hydrogenase H-cluster, a [4Fe-4S] cubane bridged by a cysteine to a 2Fe subcluster containing CO, CN Ϫ , and dithiomethylamine ligands (Figs. 1B and 2B) (3). The reversible conversion of methylenetetrahydromethanopterin to methenyltetrahydromethanopterin and H 2 is catalyzed at a mononuclear iron active site, the ligands of which include two CO factors and one guanylylpyridinol cofactor (Figs. 1C and 2C) (4).Metal cofactor assembly begins with the synthesis of FeS clusters by either the housekeeping iron-sulfur cluster assembly machinery (5) in the case of [FeFe]-hydrogenase or the Nif (nitrogen fixation)-specific homologs in the case of nitrogenase. The biosynthetic steps necessary for preparation of [FeFe]-hydrogenase and nitrogenase active sites require additional dedicated proteins that include scaffolds for assembly of the nascent cluster, as well as radical S-adenosyl-L-methionine (SAM) 2 enzymes that synthesize unique non-protein ligands. Whereas less is known about the assembly of the iron-guanylylpyridinol (FeGP) cofactor of the [Fe]-hydrogenase, it is clear that radical SAM chemistry is involved (6).It is interesting to note that in all three enzymes discussed here, the metal cofactors are organometallic in nature and exist with only minimal protein coordination (Figs. 1 and 2). The FeMo cofactor of nitrogenase is covalently bound to the protein by only one histidine and one cysteine ligand; the 2Fe subcluster of the H-cluster of [FeFe]-hydrogenase and the FeGP cofactor of [Fe]-hydrogenase have only a single cysteine ligand.Radical SAM chemistry is required for the synthesis of each of these inorganic cofactors, despite the fact that there is no single non-protein ligand shared by all three cofactors. The requirement for radical SAM chemistry to synthesize this diverse array of non-protein ligands reflects the wide variety of chemical transformations that can be initiated by hydrogen atom abstraction (7). Nitrogenase FeMo Cofactor BiosynthesisFound in a variety of bacteria and some methanogenic archaea, nitrogenase plays a critical role in the global nitrogen cycle by reducing N 2 to NH 3 (Fig. 1A). Cleavage of the triple bond in N 2 requires considerable energy, as reflected in the high temperatures, high pressures, and catalysts required to attain practical rates of N 2 reduction industrially (8). In contrast, nitrogenase employs ATP to drive the process at atmospheric pressure and ambient tempe...

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“…HydE is a member of the RS family and is essential for [FeFe]-hydrogenase maturation [6,29]. Several years ago, structures of TmHydE were reported from a crystal form that diffracted to high resolution (1.25 Å) [30].…”

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

confidence: 99%

“…Although many unresolved questions remain, using a biosynthetic cocktail (including maturases, substrates and cofactors) to achieve efficient transfer of labelled 57 Fe from HydG to the [2Fe] H subcluster of HydA [16] supports model (ii). Direct interactions between the maturase proteins HydE and HydG with HydF have been demonstrated [29,39] and GTP has been shown to accelerate the dissociation of HydE:HydF and HydG:HydF complexes [40], although the significance of these interactions and the GTPase activity awaits more detailed functional analysis.…”

Section: Hydf As a Scaffold For Hyda Maturationmentioning

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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Biochemical Analysis of the Interactions between the Proteins Involved in the [FeFe]-Hydrogenase Maturation Process

Cited by 36 publications

References 25 publications

Identification of Global Ferredoxin Interaction Networks in Chlamydomonas reinhardtii

Identification of Global Ferredoxin Interaction Networks in Chlamydomonas reinhardtii

Radical S-Adenosyl-l-methionine Chemistry in the Synthesis of Hydrogenase and Nitrogenase Metal Cofactors

Metallocofactor assembly for [FeFe]-hydrogenases

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