Metals, also homeostasis

In addition to the reduction potentials of different terminal electron acceptors utilized in the anaerobic ETC, presence of other compounds has also found to be essential for the efficiency of anaerobic respiration. One important factor is metal ion homeostasis. This is because many enzymes in the anaerobic respiratory pathways are metal containing enzymes. It was reported that expression of operons containing enzymes in the anaerobic respiration pathway is highly reduced upon iron limitation under anaerobic conditions while the aerobic respiration pathways are only modestly affected in E. coli (Cotter et al. 1992). Moreover, induction of DMSO respiration pathways is reported to be dependent on the molybdate uptake (McNichoIas et al. 1998). 

In mammals, as in yeast, several different metallothionein isoforms are known, each with a particular tissue distribution (Vasak and Hasler, 2000). Their synthesis is regulated at the level of transcription not only by copper (as well as the other divalent metal ions cadmium, mercury and zinc) but also by hormones, notably steroid hormones, that affect cellular differentiation. Intracellular copper accumulates in metallothionein in copper overload diseases, such as Wilson s disease, forming two distinct molecular forms one with 12 Cu(I) equivalents bound, in which all 20 thiolate ligands of the protein participate in metal binding the other with eight Cu(I)/ metallothionein a molecules, with between 12-14 cysteines involved in Cu(I) coordination (Pountney et ah, 1994). Although the role of specific metallothionein isoforms in zinc homeostasis and apoptosis is established, its primary function in copper metabolism remains enigmatic (Vasak and Hasler, 2000). 

Iron chelators can also be used to selectively bind iron in areas where oxidative stress is observed, thereby preventing the iron from taking part in Fenton reactions without interfering with normal iron homeostasis. Charkoudian et al. have developed boronic acid and boronic ester masked prochelators, which do not bind metals unless exposed to hydrogen peroxide (237,238). The binding of these chelators to iron(III) prevents redox cycling. Similar studies of these systems have been performed by a separate group (239,240). 

In many crucial biological processes, such as oxygen transport, electron transport, intermediary metabolism, metals play an important part. Therefore, disorders of metal homeostasis, metal bioavailability or toxicity caused by metal excess, are responsible for a large number of human diseases. We have already mentioned disorders of iron metabolism (see Chapter 7) and of copper metabolism (see Chapter 14). The important role, particularly of redox metals such as copper and iron, and also of zinc, in neurodegenerative diseases, such as Parkinson s disease, Alzheimer s disease, etc. has also been discussed (see Chapter 18). We will not further discuss them here. 

Although the S. cerevisiae MT gene is also glucose-repressible (see Butt Ecker, 1987), the primary role of this protein appears to be in metal homeostasis. This conclusion must also be drawn for the N. crassa MT, the expression of which does not appear to be regulated by anything other than Cu (Munger et al., 1987). 

Once inside the organism, organic chemicals and metals are dealt with differently. Organic chemicals generally distribute based on their chemical properties (e.g., molecular size, lipophilicity, stereochemistry) and are eliminated through metabolism (phases I and II) or excretion (for example, renal excretion in mammals) of either the parent compound or the metabolites. Metals, on the other hand, can be split into essential and nonessential elements. Biochemical mechanisms and pathways have evolved to regulate essential metals with physiological functions. However, nonessential metals due to physicochemical similarities also use some of these pathways and thus affect the homeostasis of essential metals. 

Copper ion homeostasis in prokaryotes involves Cu ion efflux and sequestration. The proteins involved in these processes are regulated in their biosynthesis by the cellular Cu ion status. The best studied bacterial Cu metalloregulation system is found in the gram-positive bacterium Enterococcus hirae. Cellular Cu levels in this bacterium control the expression of two P-type ATPases critical for Cu homeostasis (Odermatt and Solioz, 1995). The CopA ATPase functions in Cu ion uptake, whereas the CopB ATPase is a Cu(I) efflux pump (Solioz and Odermatt, 1995). The biosynthesis of both ATPases is regulated by a Cu-responsive transcription factor, CopY (Harrison et al., 2000). In low ambient Cu levels Cop Y represses transcription of the two ATPase genes. On exposure to Cu(I), CopY dissociates from promoter/operator sites on DNA with a for Cu of 20 jlM (Strausak and Solioz, 1997). Transcription of copA and copB proceeds after dissociation of CuCopY. The only other metal ions that induce CopY dissociation from DNA in vitro are Ag(I) and Cd(II), although the in vivo activation of copA and copB is specihc to Cu salts. The CuCopY complex is dimeric with two Cu(I) ions binding per monomer (C. T. Dameron, personal communication). The structural basis for the Cu-induced dissociation of CopY is unknown. Curiously, CopY is also activated in Cu-dehcient cells, but the mechanism is distinct from the described Cu-induced dissociation from DNA (Wunderh-Ye and Solioz, 1999). 

---