Mitochondria coupling

How do mitochondria couple such different types of reactions as electron transfer and the formation of a phosphate anhydride bond We explore this question in the following sections. 

Energy-linked transhydrogenase, a protein in the inner mitochondrial membrane, couples the passage of protons down the electrochemical gradient from outside to inside the mitochondrion with the transfer of H from intramitochondrial NADH to NADPH for intramitochondrial enzymes such as glutamate dehydrogenase and hydroxylases involved in steroid synthesis. 

It is important to appreciate that this principle of coupling-in-series underlies all biochemical pathways or processes, e.g. glycolysis, generation of ATP in the mitochondrion, protein synthesis from amino acids or a signal transduction pathway. Indeed, despite the fundamental importance of signalling pathways in biochemistry, a thermodynamic analysis of such a pathway has never been done, but the principles outlined above must apply even to signalling pathways. 

Oxidizible substrates from glycolysis, fatty acid or protein catabolism enter the mitochondrion in the form of acetyl-CoA, or as other intermediaries of the Krebs cycle, which resides within the mitochondrial matrix. Reducing equivalents in the form of NADH and FADH pass electrons to complex I (NADH-ubiquinone oxidore-ductase) or complex II (succinate dehydrogenase) of the electron transport chain, respectively. Electrons pass from complex I and II to complex III (ubiquinol-cyto-chrome c oxidoreductase) and then to complex IV (cytochrome c oxidase) which accumulates four electrons and then tetravalently reduces O2 to water. Protons are pumped into the inner membrane space at complexes I, II and IV and then diffuse down their concentration gradient through complex V (FoFi-ATPase), where their potential energy is captured in the form of ATP. In this way, ATP formation is coupled to electron transport and the formation of water, a process termed oxidative phosphorylation (OXPHOS). 

In eukaryotes, most of the reactions of aerobic energy metabolism occur in mitochondria. An inner membrane separates the mitochondrion into two spaces the internal matrix space and the intermembrane space. An electron-transport system in the inner membrane oxidizes NADH and succinate at the expense of 02, generating ATP in the process. The operation of the respiratory chain and its coupling to ATP synthesis can be summarized as follows ... 

Calcium levels are believed to be controlled in part at least by the uptake and release of Ca2+ from mitochondria.172"174 The capacity of mitochondria for Ca2+ seems to be more than sufficient to allow the buffering of Ca2+ at low cytosol levels. Mitochondria take up Ca2+ by an energy-dependent process either by respiration or ATP hydrolysis. It is now agreed that Ca2+ enters in response to the negative-inside membrane potential developed across the inner membrane of the mitochondrion during respiration. The uptake of Ca2+ is compensated for by extrusion of two H+ from the matrix, and is mediated by a transport protein. Previous suggestions for a Ca2+-phosphate symport are now discounted. The possible alkalization of the mitochondrial matrix is normally prevented by the influx of H+ coupled to the influx of phosphate on the H - PCV symporter (Figure 10). This explains why uptake of Ca2+ is stimulated by phosphate. Other cations can also be taken up by the same mechanism. 

Electrons in the iron-sulfur clusters of NADH-Q oxidoreduetase are shuttled to coenzyme Q. The flow of two electrons from NADH to coenzyme Q through NADH-Q oxidoreduetase leads to the pumping offour hydrogen ions out of the matrix of the mitochondrion. The details of this process remain the subject of active investigation. However, the coupled electron- proton transfer reactions of Q are crucial. NADH binds to a site on the vertical arm and transfers its electrons to FMN. These electrons flow within the vertical unit to three 4Fe-4S centers and then to a bound Q. The reduction of Q to... 

The inconsistency between experiment and prediction must lead to the rejection of the model used to describe the system. In the case of oxidative phosphorylation this has led to a refined model, in which the chemiosmotic coupling is visualized as taking place within units of one (or a few) respiratory chain(s) plus ATP synthase, while the pumped protons have only limited access to the bulk phase inside and/or outside the mitochondrion [42]. This more refined model can again be tested by deriving from it flux-force relations according to the MNET approach. A discussion of the refined model can be found in Ref. 43. 

Protoiis are extruded from the mitochondrion at the sites of NADH dehydrogenase, at the cytochrome b/c complex, and at cytochrome c oxidase. The exact number of protons driven out per electron at each of these steps has been difficult to determine. The ratio can vary under different experimental conditions. Extrusion of a proton at any particular step does not seem to be tightly coupled to the passage of an electron down the respiratory chain however, it is generally accepted that the passage of two electrons down the entire respiratory chain results in the translocation of 12 protons. 

The mitochondrial inner membrane, cristae, and matrix are the sites of most reactions involving the oxidation of pyruvate and fatty acids to CO2 and H2O and the coupled synthesis of ATP from ADP and Pj. These processes Involve many steps but can be subdivided into three groups of reactions, each of which occurs in a discrete membrane or space in the mitochondrion (Figure 8-7) ... 

In the mitochondrion, the proton-motive force is generated by coupling electron flow from NADH and FADH2 to O2 to the uphill transport of protons from the matrix across the inner membrane to the Intermembrane space. 

A FIGURE 18-1 Overview of synthesis of major membrane lipids and their movement into and out of cells. Membrane lipids (e.g., phospholipids, cholesterol) are synthesized through complex multienzyme pathways that begin with sets of water-soluble enzymes and intermediates in the cytosol (D) that are then converted by membrane-associated enzymes into water-insoluble products embedded in the membrane (B), usually at the interface between the cytosolic leaflet of the endoplasmic reticulum (ER) and the cytosol. Membrane lipids can move from the ER to other organelles (H), such as the Golgi apparatus or the mitochondrion, by either vesicle-mediated or other poorly defined mechanisms. Lipids can move into or out of cells by plasma-membrane transport proteins or by lipoproteins. Transport proteins similar to those described in Chapter 7 that move lipids (0) include sodium-coupled symporters that mediate import CD36 and SR-BI superfamily proteins that can mediate... 

During the passage through the Fo channel, and interacting with the F ATPase, the movement of the hydrogen ions into the mitochondrion are coupled with... 

The return of H+ back to the mitochondrion is coupled with ATP formation. 

Ascorbic acid is now well established as an essential factor in many hydroxylation reactions of the type RH + O ROH. On the face of it this seems a paradoxical role for a reducing substance but not if one treats the vitamin as a redox couple, e.g. ascorbic acid/dehydroascorbic acid (H2A/A) which will undergo cycling, like the cytochromes. After all, what is really meant by reference to, say, cytochrome c in the context of its role in the mitochondrion is cytochrome c (Fe )/ cytochrome c (Fe ) because, in helping to transfer electrons from metabolites towards oxygen, the cytochrome molecule continually... 

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