Structural differences between the ready and unready oxidized states of [NiFe] hydrogenases

Volbeda, Anne; Martin, Lydie; Cavazza, Christine; Matho, Michael H.; W., Faber Bart; Roseboom, Winfried; J., Albracht Simon P.; Garcin, Elsa D.; Rousset, Marc; Fontecilla‐Camps, J.C.

doi:10.1007/s00775-005-0632-x

Cited by 296 publications

(327 citation statements)

References 69 publications

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“…Cys-SOH and Cys-SO 2 H serve as catalytically essential redox centers, transient intermediates during peroxide reduction, sensors for the intracellular redox status, catalytic active sites, etc. (29,30,32,33). Compared with various proteins containing reactive cysteine residues, NHase and thiocyanate hydrolase have both post-translational oxidized Cys-SOH and Cys-SO 2 H in their active sites (8 -12, 26 -28); both cysteines play an important role in the catalytic reaction but do not play any catalytic redox role (29).…”

Section: Discussionmentioning

confidence: 99%

“…Oxidized cysteine residues Cys-SO 2 H or Cys-SOH are known to play roles in diverse processes, including signal transduction, oxygen metabolism and the oxidative stress response, transcription regulation, and metal coordination in various proteins such as NADH peroxidase (29,30), peroxiredoxins (31), hydrogenase (32,33), and so on (8 -12, 26 -28, 34). Among these enzymes, NHase and thiocyanate hydrolase are intriguing ones because they possess both Cys-SO 2 H and Cys-SOH as ligands of the metal center and neither residue plays any catalytic redox role at all (29).…”

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confidence: 99%

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Self-subunit Swapping Chaperone Needed for the Maturation of Multimeric Metalloenzyme Nitrile Hydratase by a Subunit Exchange Mechanism Also Carries Out the Oxidation of the Metal Ligand Cysteine Residues and Insertion of Cobalt

Zhou¹,

Hashimoto²,

Kobayashi

2009

Journal of Biological Chemistry

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The incorporation of cobalt into low molecular mass nitrile hydratase (L-NHase) of Rhodococcus rhodochrous J1 has been found to depend on the ␣-subunit exchange between cobalt-free L-NHase (apo-L-NHase lacking oxidized cysteine residues) and its cobalt-containing mediator (holo-NhlAE containing Cys-SO 2 ؊ and Cys-SO ؊ metal ligands), this novel mode of post-translational maturation having been named self-subunit swapping, and NhlE having been recognized as a self-subunit swapping chaperone (Zhou, Z., Hashimoto, Y., Shiraki, K., and Kobayashi, M. (2008) Proc. Natl. Acad. Sci. U. S. A. 105, 14849 -14854). We discovered here that cobalt was inserted into both the cobaltfree NhlAE (apo-NhlAE) and the cobalt-free ␣-subunit (apo-␣-subunit) in an NhlE-dependent manner in the presence of cobalt and dithiothreitol in vitro. Matrix-assisted laser desorption ionization time-of-flight mass spectroscopy analysis revealed that the non-oxidized cysteine residues in apo-NhlAE were posttranslationally oxidized after cobalt insertion. These findings suggested that NhlE has two activities, i.e. cobalt insertion and cysteine oxidation. NhlE not only functions as a self-subunit swapping chaperone but also a metallochaperone that includes a redox function. Cobalt insertion and cysteine oxidation occurred under both aerobic and anaerobic conditions when Co 3؉ was used as a cobalt donor, suggesting that the oxygen atoms in the oxidized cysteines were derived from water molecules but not from dissolved oxygen. Additionally, we isolated apo-NhlAE after the self-subunit swapping event and found that it was recycled for cobalt transfer into L-NHase.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Self-subunit Swapping Chaperone Needed for the Maturation of Multimeric Metalloenzyme Nitrile Hydratase by a Subunit Exchange Mechanism Also Carries Out the Oxidation of the Metal Ligand Cysteine Residues and Insertion of Cobalt

Zhou¹,

Hashimoto²,

Kobayashi

2009

Journal of Biological Chemistry

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“…Most [NiFe] hydrogenases, however, are reversibly inactivated by molecular oxygen (13,14). Under electron-rich conditions in the presence of O 2 a mono-oxo species, most probably a hydroxide, is formed in the bridging position between nickel and iron (15)(16)(17). On the basis of EPR spectroscopy, this paramagnetic "ready inactive" state has been designated as Ni r -B. Incubation of the enzyme with O 2 under electron-poor conditions results in the so-called "unready inactive" Ni u -A state.…”

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confidence: 99%

“…It is anticipated that a di-oxo species (e.g. hydroperoxide) binds in the bridging position (16,17). However, the nature of this ligand is still a matter of debate (18).…”

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confidence: 99%

Spectroscopic Insights into the Oxygen-tolerant Membrane-associated [NiFe] Hydrogenase of Ralstonia eutropha H16

Saggu

Zebger

Ludwig

et al. 2009

Journal of Biological Chemistry

106

209

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Hydrogenases are metalloenzymes that catalyze the reversible cleavage of H 2 into protons and electrons and play a pivotal role in the energy metabolism of many microorganisms (1). They are grouped into three phylogenetically distinct classes as follows: the di-iron [FeFe], nickel-iron [NiFe], and iron-sulfur cluster-free [Fe] hydrogenases (2-6). The basic module of [NiFe] hydrogenases consists of two subunits, a large subunit that contains the [NiFe] active site and a small subunit that accommodates one to three electron-transferring iron-sulfur

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“…[NiFe]hydrogenases are particularly interesting because of their heterobinuclear active site and have been studied extensively (3)(4)(5)(6)(7)(8)(9)(10)(11)(12). Thiolate-bridged heterobimetallic Ni-Fe complexes, therefore, have also received considerable attention because of their importance as structural, spectroscopic, and functional models for the active site of the enzyme (13)(14)(15).…”

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confidence: 99%

Modulation of the electronic structure and the Ni–Fe distance in heterobimetallic models for the active site in [NiFe]hydrogenase

Zhu

Marr

Wang

et al. 2005

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

155

154

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Reaction of the mononuclear Ni(II) thiolate complexes [Ni(L)]hydrogenase ͉ iron ͉ nickel ͉ thiolates ͉ density functional theory calculations H ydrogenases catalyze the reversible oxidation of molecular hydrogen and play a vital role in anaerobic metabolisms of a wide variety of microorganisms (1, 2).[NiFe]hydrogenases are particularly interesting because of their heterobinuclear active site and have been studied extensively (3-12). Thiolate-bridged heterobimetallic Ni-Fe complexes, therefore, have also received considerable attention because of their importance as structural, spectroscopic, and functional models for the active site of the enzyme (13-15). The structure of the oxidized inactive form of [NiFe]hydrogenase in Desulfovibrio gigas has been determined by x-ray crystallography (4, 5) and confirms an unusual heterobimetallic Ni-Fe core incorporating two terminal thiolates and two bridging thiolates at the Ni ion, with the bridging thiolates binding to a Fe(CN) 2 (CO) fragment with a Ni-Fe separation of 2.9 Å (Fig. 1). The vast majority of low-molecular-weight mimics have sought to model these structural patterns and this Ni-Fe separation (13-15).Of particular interest to us was the reported x-ray absorption study of [NiFe]hydrogenase from Chromatium vinosum that estimated the Ni-Fe separation as 2.5-2.6 Å in the reduced and active Ni-SI form of the enzyme (16). This finding was further supported by theoretical (17) and mechanistic (18, 19) studies that suggest a shorter Ni-Fe separation (2.5 Å) in the active form of the enzyme compared with that observed in the structure of its oxidized, inactive form (2.9 Å) (4). Furthermore, recent crystal structure determinations of [NiFe]hydrogenases from Desulfovibrio vulgaris Miyazaki F (8) and D. gigas (11) also exhibit shorter Ni-Fe separations (2.55 and 2.53 Å, respectively), while Cramer and coworkers (20) have identified spin-state changes brought about by stereochemical distortion and͞or addition of further ligands at the active site. We therefore were challenged to synthesize low-molecular-weight heterobinuclear Ni-Fe complexes that would mimic not only the short Ni-Fe distance of the Ni-SI, Ni-C, and related reduced centers, but also to model the proposed stereochemical distortions and spin-state changes at the Ni center on binding to Fe fragment(s). Materials and MethodsAll operations were carried out at room temperature under a pure argon atmosphere by using standard Schlenk techniques. [Ni(pdt) (dppe)] (21, 22) and [Fe(CO) 3 (BDA)] (23) (pdt, propanedithiolate; dppe, 1,2-diphenylphosphinoethane; BDA, benzylidene acetone) were prepared by methods described in the literature. Fe 2 (CO) 9 and Fe 3 (CO) 12 were purchased from Aldrich and used as received. All solvents were dried and distilled before use. CH 2 Cl 2 was distilled from CaH 2 and benzene from Na͞benzophenone ketyl under argon. All calculations on complex 3 were carried out with the program ORCA (24). The functional for the geometry optimization was BP86 (25,26), and the basis set was TZVP on lig...

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Structural differences between the ready and unready oxidized states of [NiFe] hydrogenases

Cited by 296 publications

References 69 publications

Self-subunit Swapping Chaperone Needed for the Maturation of Multimeric Metalloenzyme Nitrile Hydratase by a Subunit Exchange Mechanism Also Carries Out the Oxidation of the Metal Ligand Cysteine Residues and Insertion of Cobalt

Self-subunit Swapping Chaperone Needed for the Maturation of Multimeric Metalloenzyme Nitrile Hydratase by a Subunit Exchange Mechanism Also Carries Out the Oxidation of the Metal Ligand Cysteine Residues and Insertion of Cobalt

Spectroscopic Insights into the Oxygen-tolerant Membrane-associated [NiFe] Hydrogenase of Ralstonia eutropha H16

Modulation of the electronic structure and the Ni–Fe distance in heterobimetallic models for the active site in [NiFe]hydrogenase

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