Nitrogenase molybdenum-iron protein FeMo-cofactor

VU Fig. 28.18 The structures of the two types of cluster unit present in the nitrogenase molybdenum-iron protein isolated from Azotobacter vinelandii (a) the P-cluster in its reduced state and (b) the FeMo cofactor. Colour code Fe, green Mo, pale grey S, yellow C, grey N, blue O, red. Each non-terminated stick represents the connection of a coordinated amino acid to the protein backbone. 

The sequential electron transfer path in nitrogenase is followed first models of the Fe4S4 cluster of the iron-protein are discussed, then mimics of the P-cluster in the molybdenum-iron protein, and finally structural and functional models of the FeMo-cofactor are summarized. 

These clusters are each composed of eight iron atoms and seven sulfide ions. In the reduced form, each cluster takes the form of two 4Fe-3S partial cubes linked by a central sulfide ion. Each cluster is linked to the protein through six cysteinate residues. Electrons flow from the P cluster to the FeMo cofactor, a very unusual redox center. Because molybdenum is present in this cluster, the nitrogenase component is also called the molybdenum-iron protein (MoFe protein). The FeMo cofactor consists of two M-3Fe-3S clusters, in which molybdenum occupies the M site in one cluster and iron occupies it in the other. The two clusters are joined by three sulfide ions. The FeMo cofactor is also coordinated to a homocitrate moiety and to the a subunit through one histidine residue and one cysteinate residue. This cofactor is distinct from the molybdenum-containing cofactor found in sulfite oxidase and apparently all other molybdenum-containing enzymes except nitrogenase. 

Fig. 7. View of the FeMo-cofactor prosthetic group of the nitrogenase MoFe protein with some of the surrounding amino acid residues where ( ) represents the molybdenum coordinated to a-His-442 and homocitrate (at the top), ( ) represents the iron, interspersed with the sulfur (O) and carbon...

It is recalled that in Chapter 9, Section 2, the electrochemical behaviour of the FeMo cofactor from FeMo-nitrogenase, was reported. It possesses a heteronuclear iron-molybdenum-sulfur (MoFe7S9) cluster, which has similarities with the above discussed iron-sulfur proteins. 

Nitrogenase, which catalyzes the reduction of N2 to two molecules of NH3, has a different molybdenum -iron cofactor (FeMo-co). It can be obtained by acid denaturation of the very oxygen-labile iron-molybdenum protein of nitrogenase followed by extraction with d i methyl formamide.655,656 The coenzyme is a complex Fe-S-Mo cluster also containing homocitrate with a composition MoFe7S9-homocitrate (see Fig. 24-3). Nitrogenase and this coenzyme are considered further in Chapter 24. 

Nitrogen Fixation in Nature The nitrogenase enzyme is a two-component protein that consists of an electron-transfer Fe protein and a catalytic protein [85]. Three different nitrogenase enzymes are known, which differ primarily in the nature of the putative active site within the catalytic protein. The most common form is the MoFe protein, in which the active site for nitrogen reduction, the so-called FeMo cofactor (FeMoco), is composed of seven irons, one molybdenum, and nine sulfides... 

Figure 12 Assembly of the molybdenum nitrogenase. Pathways of FeMo cofactor biosynthesis, Fe protein maturation, and MoFe protein assembly are indicated in dotted, dashed, and solid arrows, respectively. The nif gene products that are also required for the vanadium and iron-only nitrogenase systems are indicated by ...

The best-characterized molybdenum nitrogenase comprises two metallosulfur proteins, that is, the molybdenum-iron (MoFe) protein and the iron (Fe) protein, both of which are essential for the enzymatic activity. The Fe protein is an U2 homodimer (encoded by nifH) of Mr 60 kDa. The two subunits are bridged by a [4Fe-4S] cluster and each has a MgATP binding site. The MoFe protein is an U2P2 tetramer (encoded by niJD and nifK) of Mr 220 kDa. It contains the [8Fe-7S] cluster (P cluster) that is bridged between each ap subunit pair and the [Mo-7Fe-9S-homocitrate] cluster (FeMo cofactor or FeMoco) that is located within each a snbunit. 

There are three catalytically necessary metal-sulfur clusters in iron-molybdenum nitrogenase the Fe4S4 cluster in the Fe protein (Section 8.22.2.1), the P cluster (Section 8.22.2.2), and the FeMo cofactor (Section 8.22.2.3). Each one will be discussed separately, along with relevant coordination complexes. 

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