Proton membrane potentials

An important mitochondrial membrane porter is used to move Ca2+ into the mitochondria while moving out two protons. Membrane potential is apparently the driving force. 

Other auxin-like herbicides (2,48) include the chlorobenzoic acids, eg, dicamba and chloramben, and miscellaneous compounds such as picloram, a substituted picolinic acid, and naptalam (see Table 1). Naptalam is not halogenated and is reported to function as an antiauxin, competitively blocking lAA action (199). TIBA is an antiauxin used in receptor site and other plant growth studies at the molecular level (201). Diclofop-methyl and diclofop are also potent, rapid inhibitors of auxin-stimulated response in monocots (93,94). Diclofop is reported to act as a proton ionophore, dissipating cell membrane potential and perturbing membrane functions. 

Reported values for A and ApH vary, but the membrane potential is always found to be positive outside and negative inside, and the pH is always more acidic outside and more basic inside. Taking typical values of A F = 0.18 V and ApH = 1 unit, the free energy change associated with the movement of one mole of protons from inside to outside is... 

A proton-motive force of approximately —250 mV is needed to achieve ATP synthesis. This proton-motive force, A, is composed of a membrane potential, A P, and a pH gradient, ApH (Chapter 21). The proton-motive force is defined as the free energy difference, AG, divided by S, Paraday s constant ... 

In chloroplasts, the value of AT is typically —50 to —100 mV, and the pH gradient is equivalent to about 3 pH units, so that — (2.3 i T/S ) ApH = —200 mV. This situation contrasts with the mitochondrial proton-motive force, where the membrane potential contributes relatively more to bsp than does the pH gradient. 

Three kinds of equilibrium potentials are distinguishable. A metal-ion potential exists if a metal and its ions are present in balanced phases, e.g., zinc and zinc ions at the anode of the Daniell element. A redox potential can be found if both phases exchange electrons and the electron exchange is in equilibrium for example, the normal hydrogen half-cell with an electron transfer between hydrogen and protons at the platinum electrode. In the case where a couple of different ions are present, of which only one can cross the phase boundary — a situation which may exist at a semiperme-able membrane — one obtains a so called membrane potential. Well-known examples are the sodium/potassium ion pumps in human cells. 

Table 1). Further determinants of blocking potency are the membrane potential and the state in which the sodium channel is in (resting, activated, inactivated). The tertiary amine group can be protonated giving most local... 

Figure 7. Mechanism of the proton-translocating ubiquinol cytochrome c reductase (complex III) Q cycle. There is a potential difference of up to 150 mV across the hydrophobic core of this complex (potential barrier represented by the vertical broken line). Cytochromes hour and b N are heme groups on the same peptide subunits of complex III which can transfer electrons across the hydrophobic core. The movement of two electrons provides the driving force to transfer two protons from the matrix to the cytosol. Diffusion of UQ and UQHj, which are uncharged, in the hydrophobic core, and lipid bilayer of the inner membrane is not influenced by the membrane potential (see Nicholls and Ferguson, 1992).

In resting muscle the high concentration of ADP does not decrease the proton gradient effectively and the high membrane potential slows electron transport. ADP, formed when ATP is hydrolyzed by myosin ATPase during contraction, may stimulate electron transport. However, the concentration of ATP (largely as its Mg salt) is buffered by its readily reversible formation from creatine phosphate catalyzed in the intermembrane space, and in other cell compartments, by the various isoenzymes of creatine kinase (reviewed by Walliman et al., 1992). 

In rat liver mitochondria, in state 4, the AP was estimated to be about 220 mV, with the membrane potential representing about 90% of this (Nicholls, 1974 Appendix 3). Similar values have been reported for human and rat skeletal muscle mitochondria in state 4 (Stumpf et al., 1982). The control of the rate of electron transport is not only determined by the availability of ADP, but also of Pj oxidizable substrates, and oxygen. There is evidence for futile cycling of protons in intact normal rat hepatocytes (Brand et al., 1993). Recently, Porter and Brand (1993) found a correlation between the proton permeability of the inner membrane of liver mitochondria and body size in animals from the mouse (20 g) to horses (150 kg) with a decrease in permeability with increasing weight of several-fold at a constant... 

Kasianowicz et al. [65] described the determination of the transport of niclosamide protons across lipid bilayer membranes by equilibrium dialysis, electrophoretic mobility, membrane potential, membrane conductance, and spectrophotometric... 

We now turn our attention to how the gradient of protons pumped by Complexes I, III and IV across the inner mitochondrial membrane into the intermembrane space, together with the associated membrane potential, is used to turn the molecular rotor that ensures... 

Living cells visualization of membranes, lipids, proteins, DNA, RNA, surface antigens, surface glycoconjugates membrane dynamics membrane permeability membrane potential intracellular pH cytoplasmic calcium, sodium, chloride, proton concentration redox state enzyme activities cell-cell and cell-virus interactions membrane fusion endocytosis viability, cell cycle cytotoxic activity... 

The chemi-osmotic theory of oxidative phosphorylation has been reviewed,74 a model for mitochondrial oxidative phosphorylation in which a membrane potential or proton gradient might transmit energy from an oxidation step to ATP synthesis has been proposed,76 and adenine nucleotide transport in mitochondria has been reviewed.76... 

To ensure the specificity of the signal, it will always be very helpful to test a unique feature of the carrier under investigation, such as specific inhibition, dependence on anions/cations/protons or trans-activation by another substrate. Sodium dependency, for example, can be tested by replacement with N-methyl-D-glucamine, whereas choline chloride as substitute of sodium is not recommended, since it may influence the membrane potential. In light of the recent finding that many transport proteins display an overlapping substrate spectrum, such specificity controls should receive adequate appreciation. 

Many parameters can be monitored, for example, free-ion concentrations, membrane potentials, activities of specific enzymes, rate of proton generation, transport of signaling molecules, and gene expression. 

Figure 9.8 Simple diagram of mitochondrial H -ion movement and axonal K -ion movement to establish membrane potentials across membranes. Note that H movement from the mitochondrial matrix to the outer surface of the inner membrane requires a specific proton pump that requires energy, which is transferred from electron transfer, whereas the K ion movement occurs via an ion channel with energy provided from the concentration difference of K ions on either side of the membrane (approximately 100-fold). The movement of both the protons and K ions generates a membrane potential. The potential across the membrane of the nerve axon provides the basis for nervous activity (see Chapter 14).

The transport systems of the inner mitochondrial membrane use various mechanisms. Metabolites or ions can be transported alone (uniport, U), together with a second substance (symport, S), or in exchange for another molecule (antiport. A). Active transport—i. e., transport coupled to ATP hydrolysis—does not play an important role in mitochondria. The driving force is usually the proton gradient across the inner membrane (blue star) or the general membrane potential (red star see p. 126). 

Kamo, N., Muratsugu, M., Hongoh, R. and Kobatake, Y., (1979) Membrane potential of mitochondria measured with an electrode sensitive to tetraphenyl phosphonium and relationship between proton electrochemical potential and phosphorylation potential in steady state. Journal of Membrane Biology, 49 (2), 105-121. 

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