Proton unidirectional

S0 -> Sx -> Tx transition Tt is a base of pyridine strength (pK 5), while S0 and Sj are practically non-basic (pK 0),1S9). Moreover, the site of protonation is Nl/2a for S0 andSi,butN5 forTj (Fig. 4). Hence, Hemmerich and Schmidt proposed87) that a regiospecific rearrangement of hydrogen bridges between flavin and an apoprotein environment may occur in a photo-excited flavoprotein, which would induce a unidirectional proton transfer. 

Since the equilibrium quotient K is small, a nonnueleophilic base is added to the reaction mixture to react with liberated protons and drive the reaction to completion (left to right). Using an excess of ROH then ensures simple unidirectional pseudo first-order (rate constant Atj) kinetics ... 

Several questions arise from these considerations. First, it is by no means clear how and why unidirectional rotation is achieved in the natural system. A hint has been given above. The proton channel is helical and is thus chiral. If one takes into account that a clockwise rotation is nothing other than a mirror image of a counterclockwise rotation, it becomes obvious why chirality is of such great importance. 

Figure 3-14. Hypothetical structures indicating possible mechanisms for transporters and channels in cell membrane (shaded region) (a) mobile carrier or porter acting as a symporter for protons (H+) and some tr ansported solute (5) (b) series of binding sites in a channel across a membrane, acting as a symporter for H+ and S (c) sequential conformations of a channel, leading to unidirectional movement of solute and (d) a protein-lined pore with multiple solute or water molecules hr single file, the most accepted version of ion or water (aquaporirr) channels.

Figure 5-19. Schematic representation of reactions occurring at the photosystems and certain electron transfer components, emphasizing the vectorial or unidirectional flows developed in the thylakoids of a chloroplast. Outwardly directed election movements occur in the two photosystems (PS I and PS II), where the election donors are on the inner side of the membrane and the election acceptors are on the outer side. Light-harvesting complexes (LHC) act as antennae for these photosystems. The plastoquinone pool (PQ) and the Cyt b(f complex occur in the membrane, whereas plastocyanin (PC) occurs on the lumen side and ferredoxin-NADP+ oxidoreductase (FNR), which catalyzes electron flow from ferredoxin (FD) to NADP+, occurs on the stromal side of the thylakoids. Protons (H+) are produced in the lumen by the oxidation of water and also are transported into the lumen accompanying electron (e ) movement along the electron transfer chain.

In the previous chapter we indicated that the components involved with electron flow are situated in the lamellar membranes of chloroplasts such that they lead to a vectorial or unidirectional movement of electrons and protons (see Fig. 5-19). We now return to this theme and focus on the gradients in H+ (protons) thus created. In the light, the difference in the chemical potential of H+ from the inside to the outside of a thylakoid acts as the energy source to drive photophosphorylation. This was first clearly recognized in the 1960s by Peter Mitchell, who received the 1978 Nobel Prize in chemistry for his enunciation of what has become known as the chemiosmotic hypothesis for interpreting the relationship among electron flow, proton movements, and ATP formation. 

Theoretical models for the functioning of bacteriorhodopsin must include entry and leaving sites for protons on the two sides of the membrane, a proton conduction pathway, and the unidirectional translocation of protons across a potential barrier somewhere inside the protein so as to accomplish net transport against an electro-... 

HPTS is a pH-sensitive fluorophore (pk, 7.3) [6]. The opposite pH sensitivity of the two excitation maxima permits the ratiometric (i.e. unambiguous) detection of pH changes in double-channel fluorescence measurements. The activity of synthetic ion channels is determined in the HPTS assay by following the collapse of an applied pH gradient. In response to an external base pulse, a synthetic ion channel can accelerate intravesicular pH increase by facilitating either proton efflux or OH influx (Fig. 11.5c). These transmembrane charge translocations require compensation by either cation influx for proton efflux or anion efflux for OH influx, i.e. cation or anion antiport (Fig. 11.5a). Unidirectional ion parr movement is osmotically disfavored (i.e. OH /M or X /H symport). HPTS efflux is possible with pores only (compare Fig. 11.5b/c). Modified HPTS assays to detect endovesiculation (Fig. 11.1c) [16], artificial photosynthesis [17] and catalysis by pores [18] exist. 

In the present case, where the experimentally determined barrier (AG ) to rotation is approximately 24.5 kcal mol-1 [44], then at 160 °C a single rotamer has a half life of about 0.17 s. Thus, if one selectively polarizes one of the three Ha, Hb and Hc (see 46 in Fig. 17) protons and - after appropriate time delays -assays for the location of that polarization, a clearcut distinction between predominantly unidirectional rotation and bidirectional rotation is available. In the former case, a disproportionate share of the polarization that has moved should be transferred to a resonance for only one of the two other protons. In the latter case, the polarization that moves should be transferred equally to the remaining two resonances. 

Assuming homogenous proton recombination and unidirectional dissociation reaction one has, for the excited-state equilibrium constant ... 

Aldol condensation between purely aliphatic aldehydes is unidirectional only between two molecules of the same aldehyde, one of which acts as the component with acidic CH whilst the other acts as proton acceptor, e.g. ... 

The protons are released to one side of an otherwise generally proton-impermeable inner mitochondrial membrane to collect the protons in the space between the inner and outer membranes of the mitochondrion. The resulting proton concentration gradient then drives formation of ATP by the quintessential protein-based machine, ATP synthase, as the protons flow back through the inner mitochondrial membrane by means of another special path effecting proton permeability. Thus there are two fundamental questions. The first is, how does electron flow within the membrane achieve unidirectional proton flow across the membrane The second is, how does the return flow of protons result in the formation of ATP, the energy coin of biology ... 

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