Proton extrusion

It is conventional to discuss the stoichiometry for proton extrusion as HV2e ratios, although there are two-, one-, and four-electron reductions at different stages in the respiratory chain. Most textbooks still assert that the flow of two electrons... 

Reuveni, M., Colombo, R., Lerner, H.-R., Pradet, A. Polkajoff-Mayber, A. (1987). Osmotically induced proton extrusion from carrot cells in suspension culture. Plant Physiology, 85, 383-8. 

Especially in dicotyledonous plant species such as tomato, chickpea, and white lupin (82,111), with a high cation/anion uptake ratio, PEPC-mediated biosynthesis of carboxylates may also be linked to excessive net uptake of cations due to inhibition of uptake and assimilation of nitrate under P-deficient conditions (Fig. 5) (17,111,115). Excess uptake of cations is balanced by enhanced net re-lea,se of protons (82,111,116), provided by increased bio.synthesis of organic acids via PEPC as a constituent of the intracellular pH-stat mechanism (117). In these plants, P deficiency-mediated proton extrusion leads to rhizosphere acidification, which can contribute to the. solubilization of acid soluble Ca phosphates in calcareous soils (Fig. 5) (34,118,119). In some species (e.g., chickpea, white lupin, oil-seed rape, buckwheat), the enhanced net release of protons is associated with increased exudation of carboxylates, whereas in tomato, carboxylate exudation was negligible despite intense proton extrusion (82,120). 

Based on the above observation, it can conceivably be concluded that the presence of Fe-WEHS-type complexes can be of relevance to the Fe nutrition of both strategy I and strategy II plants (Fig. 1). In the case of strategy I plants, an effect on the mechanisms of active proton extrusion should also be considered (see Sect. V). 

As we saw in the previous section, Strategy 1 plants utilize ferric reductases, with NADPH as electron donor, coupled to proton extrusion and a specific Fe(II) transport system localized in the root plasma membrane. Saccharomyces cerevisiae also uses cell surface reductases to reduce ferric iron, and in early studies (Lesuisse et ah, 1987 ... 

Cells drive active transport in a variety of ways. The plasma-membrane Na+-K+ pump of animal cells (a) and the plasma-membrane H+ pump of anaerobic bacteria (b) are driven by the hydrolysis of ATP. Eukaryotic cells couple the uptake of neutral amino acids to the inward flow of Na+ (c). Uptake of /3-gal actosidcs by some bacteria is coupled to inward flow of protons (d). Electron-transfer reactions drive proton extrusion from mitochondria and aerobic bacteria (e). In halophilic bacteria, bacteriorhodopsin uses the energy of sunlight to pump protons (/). E. coli and some other bacteria phosphorylate glucose as it moves into the cell and thus couple the transport to hydrolysis of phosphoenolpyruvate (g). 

Mechanisms that then restore the basal cytosolic [Ca2+] levels remain unclear. Besides a Ca2+-ATPase on the osteoclast dorsal surface relatively little is known of alternative or parallel methods for Ca2+ extrusion (Zaidi et al., 1993) although there is recent functional, evidence for a Na+/Ca2+ exchanger that, in analogy to the regulation of cytoplasmic [Ca2+] in cardiac muscle could be linked to the proton extrusion that is a primary determinant of the rate and extent of bone resorption (Moonga et al., 2001). 

TABLE 9.1. Effect of Different Humic Substances Fractions (HS) on Active Proton Extrusion from Intact Roots and on H+-ATPase Activty of Plasma Membrane (PM) Vesicles Isolated from Different Plant Roots... 

Cocucci, R. DeMichelis, M. I. Regulation of proton extrusion by plant hormones and cell elongation. Phusiol. Veg., 1975, 13(4), 797-811. 

The absence of ADP is acting, in effect, as an inhibitor of electron transport, for reasons discussed in Prob. 14.6 below. Hence, by application of the crossover theorem (Chap. 10), there are large differences in the reduction of sites of the electron-transport-chain between NAD and coenzyme Q, between cytochrome b and cytochrome c, and between cytochrome c and cytochrome a. Therefore, the absence of ADP must be inhibiting electron transport at these points in fact, these are the sites of proton extrusion leading to ATP synthesis during electron transport. 

After discussing the generation and quantitation of the potential term of the proton circuit, we shall now turn to the proton current, and examine the factors which control the flux of protons around the circuit. Although it is not possible to determine the proton current under steady-state conditions directly, the parameter may be calculated indirectly from the respiratory rate and the stoicheiometry of proton extrusion by the respiratory chain. It is outside the scope of this chapter to discuss the contentious issue of the proton stoicheiometries of the complexes, but the important feature is that, unless the complexities of variable stoicheiometry are invoked, respiration and proton current vary in parallel. 

This net proton extrusion results in a net acidification of the external medium and an alkalinization of the matrix (Fig. 2.5), and should be distinguished from the steady-state cycling which occurs during the operation of the proton circuit for ATP synthesis. Clear, alkalinization of the matrix cannot continue indefinitely, and the limitation which is set is essentially thermodynamic - the respiratory chain is incapable of maintaining a proton electrochemical potential in excess of 200-230... 

In the thermogenic plant mitochondria the heat evolution has been claimed to be accomphshed by a non-energy conserving system (alternate oxidase system) which is not coupled to proton extrusion and ADP phosphorylation (for review see Ref. 1). 

Within the last decade we have obtained a tentative concept of the molecular basis for this mammalian mitochondrial thermogenesis, and we know that in contrast to the thermogenic plant mitochondria, substrate oxidation in brown adipose tissue mitochondria is basically energy conserving, with proton extrusion occurring [5], with respiratory control, and with an ability, in principle, to capture the chemical energy in the form of ATP. 

The essential role of cytochrome c release from injured mitochondria in the activation of caspase 9 has been alluded to above. This pathway is especially important in proapoptotic stimuli that are not initiated by surface receptors for apoptosis, such as UV irradiation, and may involve mitochondrial dependent pathways [83]. Continued respiration in the presence of an open mitochondrial pore may result in the generation of reactive oxygen species. Release of cytochrome c may be mediated by the opening of the mitochondrial FT pore, a non-selective channel whose composition is only partially defined [84]. Inhibitors of FT pore opening, such as cyclosporine, which binds to the adenine nucleotide translocator (ANT), a component of the FT pore, and bongkrekic acid, as well as Bcl-2, prevent cytochrome c release and inhibit apoptosis [85] whereas activators of the FT pore, such as atractyloside and Bax induce it [86]. Oxidants can rupture the outer membrane of mitochondria and release caspase-activating proteins [87]. Some studies have shown cytochrome c release before collapse of the mitochondrial membrane potential [83] suggesting alternate control of the FT pore. Many, but not all, of the members of the Bd-2 family of proteins reside in the inner mitochondrial membrane, form ionic channels in hpid membranes and increase rates of proton extrusion in mitochondria [88] and thus may control the FT pore. The antiapoptotic and mitochondrial affects of Bd-2 are independent of caspase activity as they occur in the presence of caspase inhibitors and also in yeast that lack caspases [86]. 

Table IV. Effect of EpiBR on Growth of Maize Root Segments v Medium pH, Proton Extrusion and Membrane Potential...

Treatment Mean % Increase in Length3 a A pH a Proton Extrusion (pmol/g FW) Membrane Potential13 (mV)... 

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