Cellular Oxidative phosphorylation

Clofibrate causes a necrotizing myopathy, particularly in patients with renal failure, nephrotic syndrome or hypothyroidism. The myopathy is painful and myokymia of unknown origin is sometimes present. The mechanism of damage is not known, but p-chlorophenol is a major metabolite of clofibrate and p-chlorophe-nol is a particularly potent uncoupler of cellular oxidative phosphorylation and disrupts the fluidity of lipid membranes. Muscle damage is repaired rapidly on the cessation of treatment. 

Van der Meer et al. [59] have described the regulation of mitochondrial respiration on the basis of the principles of irreversible thermodynamics. According to these authors, and thus in contrast to the view of Holian et al. [53], the entire system of oxidative phosphorylation in the hepatocyte is displaced from equilibrium. They observed a linear relationship between the rate of oxygen consumption and the affinity of the entire oxidative pho.sphorylation system, a term which includes the cytosolic phosphate potential, the mitochondrial NADH/NAD ratio and the partial pressure of oxygen. They concluded that the adenine nucleotide translocator may be a rate-limiting step in cellular oxidative phosphorylation. 

The processes of electron transport and oxidative phosphorylation are membrane-associated. Bacteria are the simplest life form, and bacterial cells typically consist of a single cellular compartment surrounded by a plasma membrane and a more rigid cell wall. In such a system, the conversion of energy from NADH and [FADHg] to the energy of ATP via electron transport and oxidative phosphorylation is carried out at (and across) the plasma membrane. In eukaryotic cells, electron transport and oxidative phosphorylation are localized in mitochondria, which are also the sites of TCA cycle activity and (as we shall see in Chapter 24) fatty acid oxidation. Mammalian cells contain from 800 to 2500 mitochondria other types of cells may have as few as one or two or as many as half a million mitochondria. Human erythrocytes, whose purpose is simply to transport oxygen to tissues, contain no mitochondria at all. The typical mitochondrion is about 0.5 0.3 microns in diameter and from 0.5 micron to several microns long its overall shape is sensitive to metabolic conditions in the cell. 

The first is cell injury (cytotoxicity), which can be severe enough to result in cell death. There are many mechanisms by which xenobiotics injure cells. The one considered here is covalent binding to cell macromol-ecules of reactive species of xenobiotics produced by metabolism. These macromolecular targets include DNA, RNA, and protein. If the macromolecule to which the reactive xenobiotic binds is essential for short-term cell survival, eg, a protein or enzyme involved in some critical cellular function such as oxidative phosphorylation or regulation of the permeability of the plasma membrane, then severe effects on cellular function could become evident quite rapidly. 

Allelopathic inhibition of mineral uptake results from alteration of cellular membrane functions in plant roots. Evidence that allelochemicals alter mineral absorption comes from studies showing changes in mineral concentration in plants that were grown in association with other plants, with debris from other plants, with leachates from other plants, or with specific allelochemicals. More conclusive experiments have shown that specific allelochemicals (phenolic acids and flavonoids) inhibit mineral absorption by excised plant roots. The physiological mechanism of action of these allelochemicals involves the disruption of normal membrane functions in plant cells. These allelochemicals can depolarize the electrical potential difference across membranes, a primary driving force for active absorption of mineral ions. Allelochemicals can also decrease the ATP content of cells by inhibiting electron transport and oxidative phosphorylation, which are two functions of mitochondrial membranes. In addition, allelochemicals can alter the permeability of membranes to mineral ions. Thus, lipophilic allelochemicals can alter mineral absorption by several mechanisms as the chemicals partition into or move through cellular membranes. Which mechanism predominates may depend upon the particular allelochemical, its concentration, and environmental conditions (especially pH). 

Not all the cellular DNA is in the nucleus some is found in the mitochondria. In addition, mitochondria contain RNA as well as several enzymes used for protein synthesis. Interestingly, mitochond-rial RNA and DNA bear a closer resemblance to the nucleic acid of bacterial cells than they do to animal cells. For example, the rather small DNA molecule of the mitochondrion is circular and does not form nucleosomes. Its information is contained in approximately 16,500 nucleotides that func-tion in the synthesis of two ribosomal and 22 transfer RNAs (tRNAs). In addition, mitochondrial DNA codes for the synthesis of 13 proteins, all components of the respiratory chain and the oxidative phosphorylation system. Still, mitochondrial DNA does not contain sufficient information for the synthesis of all mitochondrial proteins most are coded by nuclear genes. Most mitochondrial proteins are synthesized in the cytosol from nuclear-derived messenger RNAs (mRNAs) and then transported into the mito-chondria, where they contribute to both the structural and the functional elements of this organelle. Because mitochondria are inherited cytoplasmically, an individual does not necessarily receive mitochondrial nucleic acid equally from each parent. In fact, mito-chondria are inherited maternally. 

However, the activation of IRP-1 by H2O2 and NO differs considerably. NO requires up to 15 h for complete activation of IRP-1 while H2O2 achieves maximum IRP-1 activation within 60 minutes. Such results suggest that H2O2 activation does not simply result from a direct oxidative attack on IRP-1 but involves additional cellular activities (phosphorylation) for IRP-1 activation. Bouton et al. (1997) suggested that the formation of peroxynitrite might be important for IRP-1 activation. However, as yet, other investigators have not confirmed this. 

Endocytosis has been shown to be dependent on metabolic activity. A block of metabolic activity might also be used to differentiate endocytosis from fusion or cellular association. Either glycolysis or oxidative phosphorylation can be affected, or both. 

Mitochondria, which are cytoplasmic organelles involved in cellular respiration, have their own chromosome, which contains 16,569 DNA base pairs (bp) arranged in a drcalar molecule. This DNA encodes 13 proteins that are subunits of complexes in the electron transport and oxidative phosphorylation processes (see Section 1, Chapter 13). In addition, mitochondrial DNA encodes 22 transfer RNAs and two ribosomal RNAs. 

In eukaryotes, the cytoplasm, representing slightly more than 50% of the cell volume, is the most important cellular compartment. It is the central reaction space of the cell. This is where many important pathways of the intermediary metabolism take place—e.g., glycolysis, the pentose phosphate pathway, the majority of gluconeogenesis, and fatty acid synthesis. Protein biosynthesis (translation see p. 250) also takes place in the cytoplasm. By contrast, fatty acid degradation, the tricarboxylic acid cycle, and oxidative phosphorylation are located in the mitochondria (see p. 210). 

Mitochondria are also described as being the cell s biochemical powerhouse, since—through oxidative phosphorylation (see p. 112)—they produce the majority of cellular ATP. Pyruvate dehydrogenase (PDH), the tricarboxylic acid cycle, p-oxidation of fatty acids, and parts of the urea cycle are located in the matrix. The respiratory chain, ATP synthesis, and enzymes involved in heme biosynthesis (see p. 192) are associated with the inner membrane. 

ATP (Adenosine 5 -triphosphate) plays a critical role in all living beings as an energy source for various enzyme activities and as a direct precursor in RNA synthesis. ATP is rapidly regenerated mainly by the glycolytic pathway and oxidative phosphorylation. A conventional luciferin-luciferase method was established a, 2), and many investigators have been using it to measure static ATP concentration (3). However it has been difficult to measure cellular ATP synthetic activity (4). Recendy, we developed... 

Integral proteins play a role in many other cellular processes. They serve as transporters and ion channels (discussed in Section 11.3) and as receptors for hormones, neurotransmitters, and growth factors (Chapter 12). They are central to oxidative phosphorylation and photosynthesis (Chapter 19) and to cell-cell and antigen-cell recognition in the immune system (Chapter 5). Integral proteins are also important players in the membrane fusion that accompanies exocytosis, en-docytosis, and the entry of many types of viruses into host cells. 

Oxidative Phosphorylation Is Regulated by Cellular Energy Needs... 

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