Nitrite reductase source

Such an involvement of an amino acid side-chain ligand switch within each catalytic cycle was a novel proposal and as such needs to be scrutinized by a variety of experimental procedures as well as analysis in the context of information known for cytochrome cd nitrite reductase from another source (see later discussion). However, it is interesting to note that something similar has been proposed for the protocate-chuate 3,4-dioxygenase enzyme from Pseudomonas putida (15). On the other hand, bacterial cytochrome c peroxidase offers an example where ligand switching seemingly relates only to an activation phenomenon. 

Blom was the first to demonstrate, in 1928, the formation of HA by an unknown mixture of bacteria which utilized nitrate as their sole nitrogen source to produce ammonia , an observation substantiated by Lindsey and Rhines who generalized this reaction to a diverse set of microorganisms capable of producing NH3 by reduction of both nitrites and nitrates. The mechanism of the 6-electron reduction of nitrite to ammonia (i.e. conversion of the [N + 02] species to by bacterial cytochrome c nitrite reductase... 

With M. gryphiswaldense, Schuler and Bauerlein (1996) recorded an Fe uptake rate from Fe " citrate of 0.86 nmol min mg dry weight and suggested that the major portion of Fe is taken up in an energy-dependent process possibly by a reductive step (Schuler, 1999). Fukumori et al. (1997) proposed that the dissimilatory nitrite reductase of M. magnetotacticum may function as an Fe" oxidizing enzyme. Later, Fuko-mori (2000) suggested an Fe "quinate complex as the source of Fe which is subsequently reduced in the cell in a microaerobic environment at about neutral pH by the iron reductase NADH (an assimilatory enzyme). 

There are a number of excellent sources of information on copper proteins notable among them is the three-volume series Copper Proteins and Copper Enzymes (Lontie, 1984). A review of the state of structural knowledge in 1985 (Adman, 1985) included only the small blue copper proteins. A brief review of extended X-ray absorption fine structure (EXAFS) work on some of these proteins appeared in 1987 (Hasnain and Garner, 1987). A number of new structures have been solved by X-ray diffraction, and the structures of azurin and plastocyanin have been extended to higher resolution. The new structures include two additional type I proteins (pseudoazurin and cucumber basic blue protein), the type III copper protein hemocyanin, and the multi-copper blue oxidase ascorbate oxidase. Results are now available on a copper-containing nitrite reductase and galactose oxidase. 

Intriguingly, the blue copper sites, especiaUy those with a carbonyl oxygen at the axial coordination position, display high affinity for Zn + ions. Mutants in which the Met is replaced by Gin or Glu preferentiaUy bind Zn + when expressed in heterologous systems, e.g., Escherichia coli. Examples include azurin, amicyanin, nitrite reductase, and possibly also plastocyanin (Diederix et al., 2000 Hibino et al., 1995 Murphy et al., 1995 Nar et al., 1992a Romero et al., 1993). In the case of azurin it has been shown that both wild-type and the Met—Gin mutant have the same affinity for both Zn +and Cu + (Romero ci a/., 1993). In addition, EXAFS studies showed that some preparations of blue copper proteins purihed from their natural sources also contain small fractions of Zn derivatives (DeBeer George, personal communication). 

Many species of bacteria also have an assimilatory nitrite reductase which is located in the cytoplasm. There is relatively little known about such enzymes but the electron donor is throught to be NADPH and the active site again has siroheme (Cole, 1988). The assimilatory nitrite reductases of both plants and bacteria use nitrite that is provided as the product of the assimilatory nitrate reductases. Nitrate is a very common natural N source for plant and bacterial growth. 

Figure 11 Nitrogen sources and metabolic pathways in marine phytoplankton. Solid circles are transporters. Boxes are the catalytic enzymes and open circles are metals associated with each enzyme. Ur, Urease NR, Nitrate Reductase NiR, Nitrite Reductase AAOx, amino acid oxidase AmOx, amine oxidase GS-GOGAT, Glutamine Synthetase- Glutamate oxy-glutarate aminotransferase (or glutamate synthase).

Direct electrochemistry has also been used (72-78) to couple the electrode reactions to enzymes for which the redox proteins act as cofactors. In the studies, the chemically reduced or oxidized enzyme was turned over through the use of a protein and its electrode reaction as the source or sink of electrons. In the first report (72, 73) of such application, the electrochemical reduction of horse heart cjd,ochrome c was coupled to the reduction of dioxygen in the presence of Pseudomonas aeruginosa nitrite reductase/cytochrome oxidase via the redox proteins, azurin and cytochrome C551. The system corresponded to an oxygen electrode in which the four-electron reduction of dioxygen was achieved relatively fast at pH 7. 

Ritchie GAF, Nicholas DJD (1972) Identification of sources of nitrous oxide produced by oxidative and reductive processes in Nitrosomonas europaea. Biochem J 126 1189-1191 Ritchie GAF, Nicholas DJD (1974) The partial characterization of purified nitrite reductase and hydroxylamine oxidase from Nitrosomonas europaea. Biochem J 138 471 180... 

Regardless of whether the process of nitrate reduction is located in photosynthetic or nonphotosynthetic tissues, it still involves a cytoplasmically located nitrate reductase [reaction (6)] and a nitrite reductase complex [reaction (7)], which is located in plastids. Possible sources of reductant for these reactions have been discussed in several reviews (e.g., Lee, 1980 Abrol et al., 1983 Smirnoff and Stewart, 1985) and the conclusion reached that in heterotrophic (nonphotosynthetic) nitrate assimilation the NADH required by nitrate reductase might be derived from glycolysis, from the oxidative pentose phosphate pathway, or even from mitochondrial dehydrogenases (see I e, 1980), whereas the pentose phosphate pathway may be of singular significance in supplying NADPH for nitrite assimilation. As indicated for root tissue by Ernes et al. 

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