Iron-sulfur cluster forms

Electron Transfer Pathways. For the Ni-Fe and Fe-Fe hydrogenases, the iron-sulfur clusters form an electron transfer chain to the putative binding site for the external electron carrier (Fig. 2B, 3B), examples of which are given in Table 1. The iron-sulfur clusters are relatively close together with an edge-to-edge distance of 1.0 nm or less. Over this distance, electron transfer has been shown to be facile in proteins (30). This is sometimes described as quantum-mechanical electron tunneling, which depends on the overlap of wave functions for the two centers. This is one reason why the active site is deeply buried the protein acts as electrical insulation to prevent unwanted electron transfer with other cellular components. 

Iron Sulfur Compounds. Many molecular compounds (18—20) are known in which iron is tetrahedraHy coordinated by a combination of thiolate and sulfide donors. Of the 10 or more stmcturaHy characterized classes of Fe—S compounds, the four shown in Figure 1 are known to occur in proteins. The mononuclear iron site REPLACE occurs in the one-iron bacterial electron-transfer protein mbredoxin. The [2Fe—2S] (10) and [4Fe—4S] (12) cubane stmctures are found in the 2-, 4-, and 8-iron ferredoxins, which are also electron-transfer proteins. The [3Fe—4S] voided cubane stmcture (11) has been found in some ferredoxins and in the inactive form of aconitase, the enzyme which catalyzes the stereospecific hydration—rehydration of citrate to isocitrate in the Krebs cycle. In addition, enzymes are known that contain either other types of iron sulfur clusters or iron sulfur clusters that include other metals. Examples include nitrogenase, which reduces N2 to NH at a MoFe Sg homocitrate cluster carbon monoxide dehydrogenase, which assembles acetyl-coenzyme A (acetyl-CoA) at a FeNiS site and hydrogenases, which catalyze the reversible reduction of protons to hydrogen gas. 

Nonrepetitive but well-defined structures of this type form many important features of enzyme active sites. In some cases, a particular arrangement of coil structure providing a specific type of functional site recurs in several functionally related proteins. The peptide loop that binds iron-sulfur clusters in both ferredoxin and high potential iron protein is one example. Another is the central loop portion of the E—F hand structure that binds a calcium ion in several calcium-binding proteins, including calmodulin, carp parvalbumin, troponin C, and the intestinal calcium-binding protein. This loop, shown in Figure 6.26, connects two short a-helices. The calcium ion nestles into the pocket formed by this structure. 

In summary, it appears that the protein has to adopt the correct fold before the Rieske cluster can be inserted. The correct folding will depend on the stability of the protein the Rieske protein from the thermoacidophilic archaebacterium Sulfolobus seems to be more stable than Rieske proteins from other bacteria so that the Rieske cluster can be inserted into the soluble form of the protein during expression with the help of the chaperonins. If the protein cannot adopt the correct fold, the result will be either no cluster or a distorted iron sulfur cluster, perhaps using the two cysteines that form the disulfide bridge in correctly assembled Rieske proteins. 

EPR studies on electron transfer systems where neighboring centers are coupled by spin-spin interactions can yield useful data for analyzing the electron transfer kinetics. In the framework of the Condon approximation, the electron transfer rate constant predicted by electron transfer theories can be expressed as the product of an electronic factor Tab by a nuclear factor that depends explicitly on temperature (258). On the one hand, since iron-sulfur clusters are spatially extended redox centers, the electronic factor strongly depends on how the various sites of the cluster are affected by the variation in the electronic structure between the oxidized and reduced forms. Theoret-... 

FIGURE 3.10 EPR of ferricyanide. Potassium ferricyanide is a general-purpose oxidant of metalloproteins. The low-spin 3c/5 Fe(III) of frozen dissolved K3Fe(CN)6 gives a broad 5=1/2 spectrum with main peak at g = 2.68 (v = 9407 MHz T = 15.5 K). The sharper feature around g = 2 is from an iron-sulfur cluster anaerobically oxidized in this experiment by ferricyanide to the [3Fe-4S]1+ form with S = 1/2. 

The fourth state with [Fe4S4]° shown in Table 6.1 was recently described as the most reduced form possible for the Fe-protein s [Fe4S4] cluster.16 Usually, only two oxidation states for a given metal-sulfur cluster are stable. Therefore a stable [Fe4S4]° state in Fe-protein s iron-sulfur cluster (as appears likely from experimental evidence presented in reference 16) would be unique because the cluster would then have three stable oxidation states, [Fe4S4]2+/1+/0. It appears also that the all-ferrous state is only stable in the protein-bound cluster and not for model... 

This may be the reason why silicon is essential, namely that it keeps aluminium in a non-toxic form as aluminium silicate. While silicon is required as a trace element in most animals, in plants, particularly grasses, and in many unicellular organisms, such as diatoms4, it is a major structural element. The importance of phosphorus and sulfur is obvious, the latter often associated with iron in an important family of proteins that contains iron-sulfur clusters. 

There are many other proteins that contain iron in a form that is neither in haem nor in iron-sulfur clusters. We have already encountered the iron storage and transport proteins, ferritin and transferrin (see Chapter 8). We propose to discuss here two other classes of iron-containing proteins, those with mononuclear non-haem iron centres and those with dinuclear non-haem iron centres. 

Zirngibl, C., van Dongen, W., Schworer, B., von Biinau, R., Richter, M., Klein, A., Thauer, R. K. (1992) H2-forming methylenetetrahydromethanopterin dehydrogenase, a novel type of hydrogenase with iron-sulfur clusters in methanogenic archaea. Eur. J. Biochem. 208, 511-20. 

Iron (Fe) is quantitatively the most important trace element (see p. 362). The human body contains 4-5 g iron, which is almost exclusively present in protein-bound form. Approximately three-quarters of the total amount is found in heme proteins (see pp. 106,192), mainly hemoglobin and myoglobin. About 1% of the iron is bound in iron-sulfur clusters (see p. 106), which function as cofactors in the respiratory chain, in photosynthesis, and in other redox chains. The remainder consists of iron in transport and storage proteins (transferrin, ferritin see B). 

A brief historical note on the structure of the iron-sulfur clusters in ferredoxins is relevant. After the first analytical results revealed the presence of (nearly) equimolar iron and acid-labile sulfur, it was clear that the metal center in ferredoxins did not resemble any previously characterized cofactor type. The early proposals for the Fe S center structure were based on a linear chain of iron atoms coordinated by bridging cysteines and inorganic sulfur (Blomstrom et al., 1964 Rabino-witz, 1971). While the later crystallographic analyses of HiPIP, PaFd, and model compounds (Herskovitz et al., 1972) demonstrated the cubane-type structure of the 4Fe 4S cluster, the original proposals have turned out to be somewhat prophetic. Linear chains of sulfide-linked irons are observed in 2Fe 2S ferredoxins and in the high-pH form of aconitase. Cysteines linked to several metal atoms are present in metallothionein. The chemistry of iron-sulfur clusters is rich and varied, and undoubtedly many other surprises await in the future. 

Nitrite reductase and sulfite reductase are enzymes found in choroplasts and in prokaryotes that reduce nitrite to ammonia and sulfite to sulfide (Scott et al., 1978). Sulfite reductase also catalyzes reduction of nitrite at a lower rate. Both enzymes contain a siroheme prosthetic group linked to an iron-sulfur cluster. In siroheme, the porphyrinoid moiety is present in the more reduced chlorin form. Because NO lies between nitrite and ammonia in oxidation state, it is a potential intermediate. 

The iron responsive element, a critical factor in the control of proteins involved in iron utilization, has been identified as the cytoplasmic form of the iron-sulfur protein aconitase (Kennedy et al., 1992). Activated macrophages have been shown to activate this element, presumably by attack of the iron-sulfur cluster by NO (Drapier et al., 1993). It has been claimed that this attack is mediated by peroxynitrite (Castro et al., 1994 Hausladen and Fridovich, 1994, but this conclusion is not universally accepted. 

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