Protein flavodoxine

Figure 2.11 Beta sheets are usuaiiy represented simply by arrows in topology diagrams that show both the direction of each (3 strand and the way the strands are connected to each other along the polypeptide chain. Such topology diagrams are here compared with more elaborate schematic diagrams for different types of (3 sheets, (a) Four strands. Antiparallel (3 sheet in one domain of the enzyme aspartate transcarbamoylase. The structure of this enzyme has been determined to 2.8 A resolution in the laboratory of William Lipscomb, Harvard University, (b) Five strands. Parallel (3 sheet in the redox protein flavodoxin, the structure of which has been determined to 1.8 A resolution in the laboratory of Martha Ludwig, University of Michigan, (c) Eight strands. Antiparallel barrel in the electron carrier plastocyanln. This Is a closed barrel where the sheet is folded such that (3 strands 2 and 8 are adjacent. The structure has been determined to 1.6 A resolution in the laboratory of Hans Freeman in Sydney, Australia. (Adapted from J. Richardson.)...

Redox catalysis is the catalysis of redox reactions and constitutes a broad area of chemistry embracing biochemistry (cytochromes, iron-sulfur proteins, copper proteins, flavodoxins and quinones), photochemical processes (energy conversion), electrochemistry (modified electrodes, organic synthesis) and chemical processes (Wacker-type reactions). It has been reviewed altogether relatively recently [2]. We will essentially review here the redox catalysis by electron reservoir complexes and give a few examples of the use of ferrocenium derivatives. 

Fig. 13.4. Binding of the ligand flavin mononucleotide (red) to the protein flavodoxine (3FXN), represented by the spatial orientation of the backbone (magenta). A terminal phosphate group (blue) of the ligand (red) orients towards the apex of a helix (yellow) of the protein backbone (magenta)...

NADPH ferredoxin —> unknown protein — flavodoxin - nitrogenase... 

Fig. 8.6 The molecular Lego approach applied to the scaffold of P450 BM3 a to generate a P450 catalytic domain electrochemically accessible through the fusion with the electron transfer protein flavodoxin b to solubilize the human membrane-bound P450 2E1 by fusion with select-...

Figure 1 The basis of comparative protein structure modeling. Comparative modeling is possible because evolution resulted in families of proteins, such as the flavodoxin family, modeled here, which share both similar sequences and 3D structures. In this illustration, the 3D structure of the flavodoxin sequence from C. crispus (target) can be modeled using other structures in the same family (templates). The tree shows the sequence similarity (percent sequence identity) and structural similarity (the percentage of the atoms that superpose within 3.8 A of each other and the RMS difference between them) among the members of the family.

In globular protein structures, it is common for one face of an a-helix to be exposed to the water solvent, with the other face toward the hydrophobic interior of the protein. The outward face of such an amphiphilic helix consists mainly of polar and charged residues, whereas the inward face contains mostly nonpolar, hydrophobic residues. A good example of such a surface helix is that of residues 153 to 166 of flavodoxin from Anabaena (Figure 6.24). Note that the helical wheel presentation of this helix readily shows that one face contains four hydrophobic residues and that the other is almost entirely polar and charged. 

However, some data have been more difficult to incorporate into the mechanism shown in Figs. 8 and 9. As reported 21) in Section II,B the Fe protein can be reduced by two electrons to the [Fe4S4]° redox state. In this state the protein is apparently capable of passing two electrons to the MoFe protein during turnover, although it is not clear whether dissociation was required between electron transfers. More critically, it has been shown that the natural reductant flavodoxin hydroquinone 107) and the artificial reductant photoexcited eosin with NADH 108) are both capable of passing electrons to the complex between the oxidized Fe protein and the reduced MoFe protein, that is, with these reductants there appears to be no necessity for the complex to dissociate. Since complex dissociation is the rate-limiting step in the Lowe-Thorneley scheme, these observations could indicate a major flaw in the scheme. 

Some aspects of the Lowe-Thomeley mechanism for nitrogenase action, which has served us well over the past 15 years, are being called into question. In particular, the necessity for protein-protein dissociation after each electron transfer, the rate-determining step with dithionite as reductant, is being questioned when the natural electron donor flavodoxin or other artificial systems are used. Some aspects of the mechanism should be reinvestigated. 

Moreover, an electron transfer chain could be reconstituted in vitro that is able to oxidize aldehydes to carboxylic acids with concomitant reduction of protons and net production of dihydrogen (213, 243). The first enzyme in this chain is an aldehyde oxidoreductase (AOR), a homodimer (100 kDa) containing one Mo cofactor (MOD) and two [2Fe—2S] centers per subunit (199). The enzyme catalytic cycle can be regenerated by transferring electrons to flavodoxin, an FMN-con-taining protein of 16 kDa (and afterwards to a multiheme cytochrome and then to hydrogenase) ... 

Reduction of Fe-protein by electron carriers such as ferredoxin or flavodoxin or by inorganic dithionite ion. 

Researchers studying the stepwise kinetics of nitrogenase electron transfer using stopped-flow kinetic techniques have presented other scenarios. One hypothesis presents kinetic evidence that dissociation of Fe-protein from MoFe-protein is not necessary for re-reduction of Fe-protein by flavodoxins.13 These authors state that the possibility of ADP-ATP exchange while Fe-protein and MoFe-protein are complexed with each other cannot be excluded and that dissociation of the complex during catalysis may not be obligatory when flavodoxin is the Fe-protein reductant. This leads to the hypothesis that MgATP binds to the preformed Fe-protein/... 

Fig. 23. Schematic drawing of the backbone of flavodoxin, a protein in which a parallel 0 sheet is the dominant structural feature. The sheet (represented by arrows) is shown from one edge, so that the characteristic twist can be seen clearly.

The doubly wound structures were first recognized as a category by Rossmann in comparing flavodoxin with lactate dehydrogenase dl. As more and more protein structures were solved which fell into this... 

Flavodoxins are a group of flavoproteins which function as electron carriers at low potential in oxidation-reduction systems. The proteins of this group contain one molecule of FMN as their prosthetic group, but, in contrast to ferredoxins, do not contain metals such as iron. 

Table 2. Molecular weight of selective flavodoxins (FMN proteins) ...

In plants, algae and cyanobacteria the light-induced charge separation of photosynthesis occurs in 2 large membrane proteins, called photosystem (PS) I and II. PS I catalyzes the ET from plastocyanin (or cytochrome c6) on the luminal side to ferrodoxin (or flavodoxin) on the stromal side of the membrane (for review see reference 177). PS I from the cyanobacterium Thermo(Y13)synechococcus (T.) elongatus was crystallized and an X-ray crystallographic structure at 2.5 A resolution has recently been obtained.18,178 Very recently, the structure from plant PS I has also been reported with a resolution of 4.4 A.179... 

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