Copper-zinc-containing SOD

SOD comprises a family of metalloproteins primarily classified into four groups copper, zinc-containing SOD (Cu, Zn-SOD), manganese-containing SOD (Mn-SOD), iron-containing SOD (Fe-SOD) and nickel-containing SOD (Ni-SOD). In the following studies, we will only focus on the uses of the former three kinds of SODs to construct SOD-based 02 biosensors since the last one, Ni-SOD, is not commercially available. 

Human cells have a manganese-containing SOD (Mn-SOD) in the mitochondria, whereas the copper- and zinc-containing SOD (Cu,Zn-SOD) is primarily present in the cytosol [39]. Two enzyme systems exist to catalyze the breakdown of H202. Firstly, the enzyme catalase, which is located in the peroxisomes, converts H202 into H20 and 02 (Eq. 2) [35]. 

Bacterial SODs typically contain either nonheme iron (FeSODs) or manganese (MnSODs) at their active sites, although bacterial copper/zinc and nickel SODs are also known (Imlay and Imlay 1996 Chung et al. 1999). Catalases are usually heme-containing enzymes that catalyze disproportionation of hydrogen peroxide to water and molecular oxygen (Eq. 10.2) (Zamocky and Koller 1999 Loewen et al. 2000). 

Although zinc itself is not redox-active, some class I enzymes containing zinc in their active sites are known. The most prominent are probably alcohol dehydrogenase and copper-zinc superoxide dismutase (Cu,Zn-SOD). AU have in common that the redox-active agent is another transition-metal ion (copper in Cu,Zn-SOD) or a cofactor such as nicotinamide adenine dinucleotide (NAD+/NADH). The Zn(II) ion affects the redox reaction only in an indirect manner, but is nevCTtheless essential and cannot be regarded simply as a structural factor. 

Antioxidant Enzymes. Reports indicate that CNS catalase activity is very poor, and only moderate amounts of GPx and SOD are present (C12, H2). Since the copper-zinc SOD gene is preferentially expressed in the neuromelanin-containing neurons within the substantia nigra, these cells may be particularly vulnerable to oxidative stress and require a high SOD content to facilitate removal of superoxide radicals. Others have also reported that SOD and GPx levels are... 

Fig. 1. Schematic overview of copper trafficking and homeostasis inside the yeast cell. The actions of Mad and Ace 1, copper-dependent metalloregulatory transcription factors, control the production of copper import [copper transporter (Ctr) and reductase (Fre)] and detoxification/sequestration [metallothionein (MT)] machineries, respectively. Three chaperone-mediated delivery pathways are shown. Atxl shuttles Cu(I) to the secretory pathway P-type ATPase Ccc2 (right). CCS delivers Cu(I) to the cytoplasmic enzyme copper-zinc superoxide dismutase (SOD) (left). Coxl7 shuttles Cu(I) to cytochrome c oxidase (CCO) in the mitochondria (bottom). Mitochondrial proteins Scol and Sco2 may also play a role in copper delivery to the CuA and CuB sites of CCO. Copper metabolism and iron metabolism are linked through the actions of Fet3, a copper-containing ferroxidase required to bring iron into the cell (lower right) (see text).

Antioxidant Functions, Both intracellular and extracellular SODs are copper- and zinc-containing enzymes, able to convert superoxide radicals to hydrogen peroxide, which can be subsequently removed by catalase and other antioxidant defenses. The plasma protein ceruloplasmin also binds copper ions and thus prevents oxidative damage from free copper ions, which can generate hydroxyl radicals. 

Superoxide dismutase (SOD) is a widely distributed enzyme that exists in a variety of forms. The copper-zinc enzyme (Cu,ZnSOD) is primarily located in the cytosol of eukaryotic cells. Mitochondria contain, in the matrix space, a distinctive cyanide-insensitive manganese-containing enzyme (MnSOD) similar to that found in prokaryotes. In addition, a ferrienzyme (FeSOD) has been identified in bacteria that is also insensitive to cyanide. Amino acid sequence homologies indicate two families of superoxide dismutases. One of these is composed of the Cu,ZnSODs and the other of MnSODs and FeSODs. All these superoxide dismutases catalyze the same reaction (2H -H O2 -h OJ H2O2 -t- O2) and with comparable efficiency. 

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