Highlights

Biological Role of Iron Complexes

Bioinorganic Reaction Mechanisms: From High-Valent Iron to Bioorganometallic Chemistry
Leonardo D. Slep and Frank Neese*
Keywords: bioinorganic chemistry · cyanide ligands · iron complexes · metalloproteins · oxygen activation

large number of enzymes contain transition-metal ions in their active center. Such enzymes efficiently catalyze a wide range of chemical reactions including those that involve activation of small molecules in a selective way and under extremely mild conditions. Unraveling the underlying structural and electronic features of these fascinating reactions is at the heart of bioinorganic chemistry. The past few years have witnessed significant progress in our understanding of many of these reactions as a result of detailed structural and spectroscopic studies as well as advanced theoretical techniques and kinetic analysis. The preparation of low-molecular-weight model complexes, although often synthetically challenging, is important to help understanding the features of a metalloprotein that are essential to its function. Moreover, the model studies may provide a chemical precedent for a hitherto unknown chemical species or postulated reaction intermediate. In a recent issue of Science two remarkable achievements in bioinorganic chemistry were reported by Münck, Nam, Que and co-workers[1] and by Böck and co-workers,[2] illustrating on how synthetic inorganic chemistry, biochemistry, and spectroscopy complement each other to answer fundamental questions in the field of metalloproteins.

A

Oxygen Activation in Nonheme Iron Enzymes
The interaction of molecular oxygen with iron in the active sites of proteins must be finely tuned to achieve either reversible O2 binding in oxygen transport (e.g. in the oxygen-carrier proteins hemoglobin and hemerythrin) or to activate O2 for further reactivity (e.g. cytochrome P450 (CYP450), mononuclear nonheme iron proteins (NHI), methane monoxygenase (MMO)). It is well known that the activation of O2 by aqueous FeII can yield a cascade of radical reactions (Fenton and Haber– Weiss reactions), which involve aggressive reactive species such as OHC, O2CÀ , or H2O2, all of which are deleterious to biological systems. The way in which

oxygen-activating iron enzymes control this hazardous chemistry and turn it constructively into specific and selective biochemical reactions has fascinated chemists for decades. The most widespread example of oxygen activation is the hemoprotein cytochrome P450CAM, which catalyzes the hydroxylation of camphor with O2 as the oxygen source. The mechanism of this reaction has been the focus of intense research efforts for more than three decades, the results of which are summarized in Scheme 1. The catalytic cycle starts with the coordination of a molecule of O2 to the active FeII site to produce an intermediate that may be considered as a FeIII–superoxide or ferrous–O2 complex. Subsequent one-electron reduction yields a FeIII–hydroper-

[*] Priv.-Doz. Dr. F. Neese, Dr. L. D. Slep Max-Planck Institut für Strahlenchemie Stiftstrasse 34–36 45470 Mülheim an der Ruhr (Germany) Fax: (+ 49) 208-306-3951 E-mail: neese@mpi-muelheim.mpg.de

Scheme 1. Important intermediates and steps in the catalytic cycle of CYP450.

2942 

2003 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

DOI: 10.1002/anie.200301649

Angew. Chem. Int. Ed. 2003, 42, 2942 – 2945

This provides a sound structural basis for future spectroscopic assignments of this structural motif and an in-depth understanding of the relationship between electronic structure. www.11-tetramethyl-1. Nam. among others. Que. OTf = CF3SO3) with iodosylbenzene (PhIO. 42. in the oxidation of Angew.813 Š). Molecular structure of the [Fe(tmc) (O)(CH3CN)]2+ dication.[7] FeIV (or even FeV) intermediates have been frequently proposed in NHI enzyme mechanisms. an oxygenatom-transfer reagent) in acetonitrile at À40 8C to yield [Fe(tmc)(O)(CH3CN)]2+ (2). formally an FeV species. Que. It has been proposed to react directly with the substrate in a fast rebound mechanism to yield the hydroxylated product and regenerate the FeIII active site. as well as by mass spectrometry. The synthesis involved the reaction of the precursor [Fe(tmc)(OTf)2] (1.8. Fascinating recent developments in the chemistry of CYP450CAM include the crystallographic characterization of several of the intermediates in the reaction cycle[3a] as well as the quantum-mechanical study of the reaction cycle with explicit inclusion of the protein environment. producing the elusive intermediate known as “compound I”. However. therefore. leading to the first spectroscopic characterization of complexes featuring the terminal [FeO]2+ and [FeN]2+ moieties[8] The article by Münck.Angewandte oxide species. lake sediments. 2003. This species was characterized by absorption. This bond is much shorter than those in bridging oxo ions in dinuclear complexes featuring FeIV (1. This is followed by heterolytic cleavage of the OÀO bond.[5] The deprotonated form.[10] and is indicative of increased p-bonding within the terminal [FeO]2+ moiety. 2942 – 2945 Chemie Scheme 2. derived from X-ray crystallography. The existence of compound I-like intermediates has been challenged on the basis of crystallographic studies which have shown that the iron centers in NHI enzymes are always coordinated to “innocent” ligands that presumably can not be ionized at the biologically relevant redox potential window. Weinheim 2943 . Its structure consists of a mononuclear iron center in a distorted octahedral environment with a short terminal FeÀO bond (1.[6] The direct reaction of low-spin FeIII–hydroperoxide complexes is. and Mössbauer spectroscopy. H2 is produced by fermentative organisms in the oxidation of sugars and other organic molecules. respectively. O22À. which do not appear to be activated for direct reaction with substrates. The short FeÀO bond length is similar to those deduced from EXAFS (extended X-ray absorption fine structure) spectroscopic measurements on high-valent porphyrins and also resembles the values based on the X-ray structural analysis of hemoproteins featuring the [FeO]2+ core. is found in side-on-bound high-spin h2FeIII–peroxo complexes. chemical modeling as well as studies on related enzymes such as catalases and peroxidases suggest that these species feature a high-valent [FeO]2+ core bound to a porphyrin radical cation rather than a genuine FeV moiety. Nam.[4] However. they provide complete structural and spectroscopic characterization of the [FeO]2+ core in a nonheme environment.[1] In this study. spectroscopy.646(3) Š. Figure 1). tmc = 1. Int. Hydrogen atoms are omitted for clarity. This has inspired synthetic chemists to attempt to prepare high-valent iron model complexes. Recent studies have shown that low-spin and high-spin FeIII hydroperoxides are activated for homolytic OÀO and FeÀO bond cleavage. The liberated electrons are transferred to protons through an electron-transport chain reaction (which involves. an alternative mechanism and has been proposed to occur. In addition. Chem. KGaA. and the intestines of animals.angewandte.[11] This remarkable results of Münck. FT IR. Ed. and co-workers provides a well-characterized model complex necessary for evaluating the feasibility of high-valent iron–oxo species in the reaction pathways of NHI enzymes.[3b] Recently.org  2003 Wiley-VCH Verlag GmbH & Co. the Fe– Figure 1. Important intermediates in the reactions of NHI enzymes. DNA by the anticancer drug bleomycin. a significant point of concern is whether the NHI enzymes could produce high-valent-iron intermediates in the absence of a porphyrin ring acting as an electron reservoir. Biosynthesis of the FeÀCN Bond in Ni–Fe Hydrogenases Hydrogen gas is an essential nutrient in virtually all forms of life that inhabit anaerobic environments.4. but have not been detected experimentally.11-tetraazacyclotetradecane. Crystallization from CH3CN/Et2O at À40 8C yielded the triflate salt 2·(OTf)2. these results allow studying the spectroscopic properties of the highvalent NHI oxo unit in the absence of impurities or other chromophores. which have proven to be as versatile as hemoproteins in oxygen-activation processes.805 Š).8. attention has been focused on mononuclear NHI proteins. and co-workers is a significant advancement towards the preparation and characterization of mononuclear high-valent nonheme iron centers. An attractive alternative to the production of highvalent intermediates is the direct reaction of FeIII–hydroperoxides to yield hydroxylated or epoxidized products (Scheme 2).4. for instance.[9] or the terminal Fe–O bond in a recently characterized complex featuring the [FeO]+ fragment (1. such as deepsea hydrothermal vent sites. The given values indicate bond lengths in Š. and reactivity in these systems. The latter behaves as an extremely powerful oxidant with a redox potential probably exceeding + 1 V.

there are two bridging cysteine fragments and an unidentified ligand (probably OHÀ).[13] The most striking structural feature of the NiFe-H2ases involves the three terminal diatomic molecules coordinated to the Fe center (Figure 2). They showed that the dehydration of S-(ndecyl)thiocarbamate assisted by ethyl polyphosphate proceeds under mild conditions and yields the corresponding thiocyanate species in a way that resem- Figure 2. which bridge to a nearby Fe center. Although several different classes of hydrogenases are known. CNÀ and CO were also found in the Scheme 3. hemoproteins involved in cell respiration). This intermediate presumably carries the SCN residue to the active site and finally transfers the electrophilic CN moiety to a nucleophilic iron center to form the Fe–CN unit. AMP = adenosine monophosphate. Pi = phosphate. they showed that organic thiocyanates are a source of CNÀ . vulgaris obtained by X-ray crystallography of the oxidized protein.angewandte.g. 42. as CO and CNÀ are not freely available in biological systems. PPi = pyrophosphate..[2] Using a combination of radio isotope labeling and mass spectrometry. Biosynthesis of the cyanide ligand in NiFe-H2ases: 1) formation of the carbamoyl adenylate on the HypF protein (F) involving an ATP–PPi exchange reaction. In active sites of other hydrogenases. 2003. Ed. Böck and co-workers explored this mechanism further with a series of elegant model experiments to probe the key steps described above. ATP = adenosine triphosphate. 2) the CONH2 group is transferred to the C-terminal cysteine of the HypE protein (E). that is. e. PDB code 2FRV. Int.[14] This result was quite surprising as both CO and CNÀ are highly toxic for biological systems because of their high affinity for the active sites of many metalloenzymes (e. 5) chemical transfer of the CN group to the active center of the NiFe-H2ase protein (LN represents the coordination sphere of the active site). a detailed spectroscopic study has provided insight into the electronic structure of the active center and of the reaction intermediates. 2942 – 2945 .Highlights S protein ferredoxin) to produce H2 gas. This final key step is catalyzed by a family of enzymes that are collectively known as hydrogenases. Chem. Weinheim www. 3) and 4) ATP-assisted dehydration involving an intermediate phosphorylation of the carbonyl oxygen atom followed by dephosphorylation to yield a thiocyanate. The CONH2 group is subsequently transferred to the thiol group of the C-terminal cysteine residue on the second protein HypE. the urea cycle) and ATP on the HypF protein. These bacteria also contain hydrogenases that catalyze the reverse reaction. The crystal structure of the NiFe-H2ase from Desulfovibrio gigas was solved in 1995[12] and showed that the active site contains a heterodinuclear NiFe site with two terminal cysteine ligands at the Ni end (Figure 2). Schematic representation of the active site of the NiFe-H2ase from D. Recently. which may be relevant to the function of H2ases. which has not been identified but is most probably an OHÀ group. The identification of CNÀ and CO ligands in the active site of metalloproteins subsequently prompted the question of how these toxic species are delivered to the active sites of the enzymes during the biosynthesis. These thiocyanates are formed as a result of the interaction of two (HypF and HypE) out of seven proteins involved in the maturation of the NiFe active site. This protein then performs a complex set of unprecedented reactions: it provides the site for dehydration of the carboxamido moiety in an ATP-dependent step to yield the cyanated cysteine (thiocyanate). The fragment X identifies the third bridging ligand. It has been proposed that this highly unusual arrangement helps to keep the Fe center in a low-spin low-valent state. KGaA.org Angew. Böck and co-workers have provided an explanation for this process in the NiFe-H2ases. ADP = adenosine diphosphate. the oxidation of H2. 2944  2003 Wiley-VCH Verlag GmbH & Co. Hydrogen gas is recycled by other organisms in the same habitat. a known intermediate in biosythetic pathways. the most common of these enzymes contain both Ni and Fe ions in their active center (NiFeH2ase). A detailed mechanism is outlined in (Scheme 3): The first step of the biosynthesis involves the formation of a carbamoyl adenylate intermediate by the reaction between carbamoyl phosphate (CP. These diatomic species were identified by FTIR spectroscopy as being one CO and two CNÀ ligands. addition.g.

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