Electrons in the Electron Transport Chain

FIGURE 11.12 Electrons transported from Complexes I and II, through the mitochondrial membrane, to Complex III (bci). Cyt c transports electrons to Complex IV (cytochrome c oxidase) outside the membrane. 

Succinate is a very simple molecule, a dicarboxylic acid where the two -COOH groups are held together by -CHj-CHj-. There is no place for an electron to localize, unless a x-system is formed. In Complex II (succinate dehydrogenase), succinate is oxidized to fumarate (HOOC-CH=CH-COOH) in a series of complicated, not fully known processes. In these processes, quinone (Q) is reduced to quinol (QH2). The electrons are transported to Complex III through the membrane. The protons are transported perpendicular to the membrane to the intermembrane space, in order to build up a proton gradient for the production of ATP. 

In Complex III, PT to the intermembrane space again takes place, driven by ET. Furthermore, the carriers are exchanged from quinones to cyt c. Cyt c moves outside the membrane and donates the electrons to Complex IV. Here the net reaction is 

The protons are again taken from the matrix and donated to the intermembrane space. 

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The tricarboxylic acid (TCA) cycle (also known as the citric acid cycle and the Krebs cycle) is a collection of biochemical reactions that oxidize certain organic molecules, generating CO2 and reducing the cofactors NAD and FAD to NADH and FADH2 [147], In turn, NADH and FADH2 donate electrons in the electron transport chain, an important component of oxidative ATP synthesis. The TCA cycle also serves to feed precursors to a number of important biosynthetic pathways, making it a critical hub in metabolism [147] for aerobic organisms. Its ubiquity and importance make it a useful example for the development of a kinetic network model. 

Coenzyme Q (CoQ) is a quinone found in the cells of all aerobic organisms. It is also called ubiquinone because it is ubiquitous (found everywhere) in nature. Its function is to carry electrons in the electron-transport chain. The oxidized form of CoQ accepts a pair of electrons from a biological reducing agent such as NADH and ultimately transfers them to O2. 

There are two general consequences to impaired functioning of the TCA cycle (1) an inability to generate ATP from fuel oxidation, and (2) an accumulation of TCA cycle precursors. For example, inhibition of pyruvate oxidation in the TCA cycle results in its reduction to lactate, which can cause a lactic acidosis. The most common situation leading to an impaired function of the TCA cycle is a relative lack of oxygen to accept electrons in the electron transport chain. 

A) Her decrease in Fe-S centers is impairing the transfer of electrons in the electron transport chain. 

Aerobic metabolism is a highly efficient way for an organism to extract energy from nutrients. In eukaryotic cells, the aerobic processes (including conversion of pyruvate to acetyl-GoA, the citric acid cycle, and electron transport) all occur in the mitochondria, while the anaerobic process, glycolysis, takes place outside the mitochondria in the cytosol. We have not yet seen any reactions in which oxygen plays a part, but in this chapter we shall discuss the role of oxygen in metabolism as the final acceptor of electrons in the electron transport chain. The reactions of the electron transport chain take place in the inner mitochondrial membrane. 

The electron in the electron transport chain is not free like in a metal wire. Therefore the electron motion in each act involves surmounting an energy barrier. As was shown in Refs. 16 and 108-110, a substantial role in this process is played by the conformations of the macromolecular components of the electron transport chain. Nevertheless, the simplest model systems of electron transport realized on bilayer lipid membranes were virtually based on the concept of a membrane as a thin liquid hydrocarbon in which a substance capable of redox transformations is dissolved, the products of this reaction being able to diffuse inside the bilayer. The electron transport from the aqueous phase containing a reducer amounts to injection of charges into the nonaqueous phase if it contains an electron acceptor ... 

The operation of the cycle requires a supply of the oxidizing agents NAD and FAD. The cycle is dependent on reactions of the electron transport chain to supply the necessary NAD and FAD (Section 13.6). Because oxygen is the final acceptor of electrons in the electron transport chain, the continued operation of the citric acid cycle depends ultimately on an adequate supply of oxygen. 

The processes in which ubiquinone participates occur in the inner mitochondrial membrane of eukaryotic cells and in the plasma membrane of bacterial cells. Ubiquinone and ubiquinol are soluble in the membrane, and they act as shuttles to transport electrons and protons from one multiprotein complex to another. Similar processes occur in the photoelectron transport chain in plants with plastiquinones as coenzymes. 

The term vitamin K2 was applied to 2-methyl-3-difarnesyl-l,4-naphthoquinone, m.p. 54 C, isolated from putrefied fish meal. It now includes a group of related natural compounds ( menaquinones ), differing in the number of isoprene units in the side chain and in their degree of unsaturation. These quinones also appear to be involved in the electron transport chain and oxidative phosphorylation. 

The ready reversibility of this reaction is essential to the role that qumones play in cellular respiration the process by which an organism uses molecular oxygen to convert Its food to carbon dioxide water and energy Electrons are not transferred directly from the substrate molecule to oxygen but instead are transferred by way of an electron trans port chain involving a succession of oxidation-reduction reactions A key component of this electron transport chain is the substance known as ubiquinone or coenzyme Q... 

The abihty of iron to exist in two stable oxidation states, ie, the ferrous, Fe ", and ferric, Fe ", states in aqueous solutions, is important to the role of iron as a biocatalyst (79) (see Iron compounds). Although the cytochromes of the electron-transport chain contain porphyrins like hemoglobin and myoglobin, the iron ions therein are involved in oxidation—reduction reactions (78). Catalase is a tetramer containing four atoms of iron peroxidase is a monomer having one atom of iron. The iron in these enzymes also undergoes oxidation and reduction (80). 

The decline in immune function may pardy depend on a deficiency of coenzyme Q, a group of closely related quinone compounds (ubiquinones) that participate in the mitochondrial electron transport chain (49). Concentrations of coenzyme Q (specifically coenzyme Q q) appear to decline with age in several organs, most notably the thymus. 

Glycolysis and the citric acid cycle (to be discussed in Chapter 20) are coupled via phosphofructokinase, because citrate, an intermediate in the citric acid cycle, is an allosteric inhibitor of phosphofructokinase. When the citric acid cycle reaches saturation, glycolysis (which feeds the citric acid cycle under aerobic conditions) slows down. The citric acid cycle directs electrons into the electron transport chain (for the purpose of ATP synthesis in oxidative phosphorylation) and also provides precursor molecules for biosynthetic pathways. Inhibition of glycolysis by citrate ensures that glucose will not be committed to these activities if the citric acid cycle is already saturated. 

FIGURE 20.1 Pyruvate produced hi glycolysis is oxidized in the tricarboxylic acid (TCA) cycle. Electrons liberated in this oxidation flow through the electron transport chain and drive the synthesis of ATP in oxidative phosphorylation. In eukaryotic cells, this overall process occurs in mitochondria. 

The TCA cycle can now be completed by converting succinate to oxaloacetate. This latter process represents a net oxidation. The TCA cycle breaks it down into (consecutively) an oxidation step, a hydration reaction, and a second oxidation step. The oxidation steps are accompanied by the reduction of an [FAD] and an NAD. The reduced coenzymes, [FADHg] and NADH, subsequently provide reducing power in the electron transport chain. (We see in Chapter 24 that virtually the same chemical strategy is used in /3-oxidation of fatty acids.)... 

Situated as it is between glycolysis and the electron transport chain, the TCA cycle must be carefully controlled by the ceil. If the cycle were permitted to run unchecked, large amounts of metabolic energy could be wasted in overproduction of reduced coenzymes and ATP conversely, if it ran too slowly, ATP would not be produced rapidly enough to satisfy the needs of the cell. Also, as just seen, the TCA cycle is an important source of precursors for biosynthetic processes and must be able to provide them as needed. 

As we have seen, the metabolic energy from oxidation of food materials—sugars, fats, and amino acids—is funneled into formation of reduced coenzymes (NADH) and reduced flavoproteins ([FADHg]). The electron transport chain reoxidizes the coenzymes, and channels the free energy obtained from these reactions into the synthesis of ATP. This reoxidation process involves the removal of both protons and electrons from the coenzymes. Electrons move from NADH and [FADHg] to molecular oxygen, Og, which is the terminal acceptor of electrons in the chain. The reoxidation of NADH,... 

Although electrons move from more negative to more positive reduction potentials in the electron transport chain, it should be emphasized that the electron carriers do not operate in a simple linear sequence. This will become evident when the individual components of the electron transport chain are discussed in the following paragraphs. 

The Electron Transport Chain Can Be Isolated in Four Complexes... 

This is a crucial point because (as we will see) proton transport is coupled with ATP synthesis. Oxidation of one FADHg in the electron transport chain results in synthesis of approximately two molecules of ATP, compared with the approximately three ATPs produced by the oxidation of one NADH. Other enzymes can also supply electrons to UQ, including mitochondrial 5w-glyc-erophosphate dehydrogenase, an inner membrane-bound shuttle enzyme, and the fatty acyl-CoA dehydrogenases, three soluble matrix enzymes involved in fatty acid oxidation (Figure 21.7 also see Chapter 24). The path of electrons from succinate to UQ is shown in Figure 21.8. 

In the third complex of the electron transport chain, reduced coenzyme Q (UQHg) passes its electrons to cytochrome c via a unique redox pathway known as the Q cycle. UQ cytochrome c reductase (UQ-cyt c reductase), as this complex is known, involves three different cytochromes and an Fe-S protein. In the cytochromes of these and similar complexes, the iron atom at the center of the porphyrin ring cycles between the reduced Fe (ferrous) and oxidized Fe (ferric) states. 

Cytochrome c, like UQ is a mobile electron carrier. It associates loosely with the inner mitochondrial membrane (in the intermembrane space on the cytosolic side of the inner membrane) to acquire electrons from the Fe-S-cyt C aggregate of Complex 111, and then it migrates along the membrane surface in the reduced state, carrying electrons to cytochrome c oxidase, the fourth complex of the electron transport chain. 

It should be emphasized here that the four major complexes of the electron transport chain operate quite independently in the inner mitochondrial membrane. Each is a multiprotein aggregate maintained by numerous strong associations between peptides of the complex, but there is no evidence that the complexes associate with one another in the membrane. Measurements of the lateral diffusion rates of the four complexes, of coenzyme Q, and of cytochrome c in the inner mitochondrial membrane show that the rates differ considerably, indicating that these complexes do not move together in the membrane. Kinetic studies with reconstituted systems show that electron transport does not operate by means of connected sets of the four complexes. 

Engelhardt s experiments in 1930 led to the notion that ATP is synthesized as the result of electron transport, and, by 1940, Severo Ochoa had carried out a measurement of the P/O ratio, the number of molecules of ATP generated per atom of oxygen consumed in the electron transport chain. Because two electrons are transferred down the chain per oxygen atom reduced, the P/O ratio also reflects the ratio of ATPs synthesized per pair of electrons consumed. After many tedious and careful measurements, scientists decided that the P/O ratio was 3 for NADH oxidation and 2 for succinate (that is, [FADHg]) oxidation. Electron flow and ATP synthesis are very tightly coupled in the sense that, in normal mitochondria, neither occurs without the other. 

When Mitchell first described his chemiosmotic hypothesis in 1961, little evidence existed to support it, and it was met with considerable skepticism by the scientific community. Eventually, however, considerable evidence accumulated to support this model. It is now clear that the electron transport chain generates a proton gradient, and careful measurements have shown that ATP is synthesized when a pH gradient is applied to mitochondria that cannot carry out electron transport. Even more relevant is a simple but crucial experiment reported in 1974 by Efraim Racker and Walther Stoeckenius, which provided specific confirmation of the Mitchell hypothesis. In this experiment, the bovine mitochondrial ATP synthasereconstituted in simple lipid vesicles with bac-teriorhodopsin, a light-driven proton pump from Halobaeterium halobium. As shown in Eigure 21.28, upon illumination, bacteriorhodopsin pumped protons... 

The second electron shuttle system, called the malate-aspartate shuttle, is shown in Figure 21.34. Oxaloacetate is reduced in the cytosol, acquiring the electrons of NADH (which is oxidized to NAD ). Malate is transported across the inner membrane, where it is reoxidized by malate dehydrogenase, converting NAD to NADH in the matrix. This mitochondrial NADH readily enters the electron transport chain. The oxaloacetate produced in this reaction cannot cross the inner membrane and must be transaminated to form aspartate, which can be transported across the membrane to the cytosolic side. Transamination in the cytosol recycles aspartate back to oxaloacetate. In contrast to the glycerol phosphate shuttle, the malate-aspartate cycle is reversible, and it operates as shown in Figure 21.34 only if the NADH/NAD ratio in the cytosol is higher than the ratio in the matrix. Because this shuttle produces NADH in the matrix, the full 2.5 ATPs per NADH are recovered. 

FIGURE 24.11 The acyl-CoA dehydrogenase reaction. The two electrons removed in this oxidation reaction are delivered to the electron transport chain in the form of reduced coenzyme Q (UQH9). 

As its name implies, the citric acid cycle is a closed loop of reactions in which the product of the hnal step (oxaloacetate) is a reactant in the first step. The intermediates are constantly regenerated and flow continuously through the cycle, which operates as long as the oxidizing coenzymes NAD+ and FAD are available. To meet this condition, the reduced coenzymes NADH and FADH2 must be reoxidized via the electron-transport chain, which in turn relies on oxygen as the ultimate electron acceptor. Thus, the cycle is dependent on the availability of oxygen and on the operation of the electron-transport chain. 

The energy released in catabolic pathways is used in the electron-transport chain to make molecules of adenosine triphosphate, ATP. ATP, the final result of food catabolism, couples to and drives many otherwise unfavorable reactions. 

Electron-transport chain (Section 29.1) The final stage of catabolism in which ATP is produced. 

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