Azurin electron transfer

Azoresorcinol, pyridyl-metal complexes dyes, 6, 74 Azurins, 6, 651, 652 copper(II) complexes, 2, 772 5, 721 electron transfer reactions, 6, 653 NMR, 6, 652 Raman spectra, 6, 652 spectra, 6, 652 thioether complexes, 2, 557 Azurite... 

Studies of ferredoxin [152] and a photosynthetic reaction center [151] have analyzed further the protein s dielectric response to electron transfer, and the protein s role in reducing the reorganization free energy so as to accelerate electron transfer [152], Different force fields were compared, including a polarizable and a non-polarizable force field [151]. One very recent study considered the effect of point mutations on the redox potential of the protein azurin [56]. Structural relaxation along the simulated reaction pathway was analyzed in detail. Similar to the Cyt c study above, several slow relaxation channels were found, which limited the ability to obtain very precise free energy estimates. Only semiquantitative values were... 

D.R. McMillin, Purdue University In addition to the charge effects discussed by Professor Sykes, I would like to add that structural effects may help determine electron transfer reactions between biological partners. A case in point is the reaction between cytochrome C551 and azurin where, in order to explain the observed kinetics, reactive and unreactive forms of azurin have been proposed to exist in solution (JL). The two forms differ with respect to the state of protonation of histidine-35 and, it is supposed, with respect to conformation as well. In fact, the lH nmr spectra shown in the Figure provide direct evidence that the nickel(II) derivative of azurin does exist in two different conformations, which interconvert slowly on the nmr time-scale, depending on the state of protonation of the His35 residue (.2) As pointed out by Silvestrini et al., such effects could play a role in coordinating the flow of electrons and protons to the terminal acceptor in vivo. 

It is interesting to speculate why nitrite reductase has its type I coppers in domains 1, whereas in hCP the mononuclear copper binding sites are retained in the domains 2,4, and 6 where they are comparatively buried in the protein. One possible reason can be related to the difference in functions of the two proteins. NR has to interact with a relatively large pseudo-azurin macromolecule in order for electron transfer to take place,... 

Simple thermodynamic considerations state that the reduction process is favoured (i.e. more positive cu(ii)/cu(p potential values are obtained) if the electron transfer is exothermic (AH° negative) and if the molecular disorder increases (AS° positive). It is therefore evident that the positive potential value for the reduction of azurin (as well as that of the most blue copper proteins) is favoured by the enthalpic factor. This means that the metal-to-ligand interactions inside the first coordination sphere (which favour the stability of the reduced form over the oxidized form) prevail over the metal complex-to-solvent interactions inside the second... 

J. W. Petrich, J. W. Longworth, and G. R. Fleming, Internal motion and electron transfer in proteins A picosecond fluorescence study of three homologous azurins. Biochemistry 26, 2711-2722 (1987). 

We may illustrate this approach to the determination of the nuclear factor by the elegant studies performed by Gray and co-workers, who have determined the thermodynamic properties and the rate temperature dependence for the electron transfer between Ru(NH3) covalently bound to the histidine residues of some proteins, and the redox eenter of these proteins [110, 111, 112, 113]. The experimental results obtained for cytochrome c [110] and azurin [111, 112] are very similar. Using the thermodynamic data and the value or the upper limit of Ea reported in these studies, we deduce from Eq. (23) ... 

Electron transfer to the protein metal center is monitored spectroscopically. In the case of a heme (FeP), a fast increase in absorbance due to direct reduction of Fe(III)P by Ru(bpy)f is followed by a slower increase in absorbance due to reduction of Fe(III)P by the Ru(II) on the protein surface. Control flash experiments with unmodified proteins show only the fast initial increase in absorbance due to Fe(III)P reduction by Ru(bpy)3. Such control experiments demonstrate for horse heart cytochrome c [21], azurin [28], and sperm whale myoglobin [14] that slow reduction of the heme by the EDTA radical produced in the scavenging step does not occur in competition with intramolecular ET. However, for Candida krusei cytochrome c, the control experiment shows evidence for slow EDTA radical reduction of the heme after initial fast reduetion by Ru(bpy)i+ [19]. 

Earlier suggestions that the two uncoordinated and invariant residues His35 (inaccessible to solvent and covered by polypeptide) and His83 (remote and 13 A from Cu) are, from effects of [H ] on rate constants (and related pKg values), sites for electron transfer may require some re-examination. Thus, it has been demonstrated in plastocyanin studies [50] that a surface protonation can influence the reduction potential at the active site, in which case its effect is transmitted to all reaction sites. In other words, an effect of protonation on rate constants need not necessarily imply that the reaction occurs at the site of protonation. His35 is thought to be involved in pH-dependent transitions between active and inactive forms of reduced azurin [53]. The proximity of... 

His35 to ligated His46 may be important in an electron transfer role or in His35 exercising some conformational control of the active site. The reduction potential of P. aeruginosa azurin increases from 300 mV (pH 8) to 360 mV (pH 5), which is believed to be related to His35 protonation. A pK of 6.6 is observed for this process, or alternatively pK s for azurin in the oxidized (6.1) and reduced (7.2) forms can be obtained [56]. 

The His35, with coordinated His46 in close proximity, has frequently been suggested as a site for electron transfer reactivity of azurin. Two processes have been detected in a temperature-jump study on the equihbration of azurin with cytochrome C551, its physiological partner [57]. The fast process is assigned to electron transfer, and the slower process to a conversion between inactive and active forms of reduced azurin. It has been concluded that the active form is protonated. A second H-bonded form of His35 is believed to result from the protonation [2]. 

More subtle factors that might affect k will be the sites structures, their relative orientation and the nature of the intervening medium. That these are important is obvious if one examines the data for the two copper proteins plastocyanin and azurin. Despite very similar separation of the redox sites and the driving force (Table 5.12), the electron transfer rate constant within plastocyanin is very much the lesser (it may be zero). See Prob. 16. In striking contrast, small oxidants are able to attach to surface patches on plastocyanin which are more favorably disposed with respect to electron transfer to and from the Cu, which is about 14 A distant. It can be assessed that internal electron transfer rate constants are =30s for Co(phen)3+, >5 x 10 s for Ru(NH3)jimid and 3.0 x 10 s for Ru(bpy)3 , Refs. 119 and 129. In the last case the excited state Ru(bpy)3 is believed to bind about 10-12 A from the Cu center. Electron transfer occurs both from this remote site as well as by attack of Ru(bpy)j+ adjacent to the Cu site. At high protein concentration, electron transfer occurs solely through the remote pathway. 

Direct electron transfer has also been achieved with many metalloproteins such as cytochrome C, horseradish peroxidase, microperoxidase (MP-11), myoglobin, hemoglobin, catalase, azurin, and so on, immobilized on different CNT-modified electrodes [45, 61, 144—153]. 

It has long been known that, under some conditions at least, electron transfer between the c and d hemes of the P aeruginosa enzyme is slow and requires times of the order of seconds (22). What does this mean It is not necessarily related to the loss of the hydroxide ligand from the d heme iron, because under some experimental conditions used, azurin (a cupredoxin) was present and the enzyme was reduced at the outset,... 

Blue copper proteins, 36 323, 377-378, see also Azurin Plastocyanin active site protonations, 36 396-398 charge, 36 398-401 classification, 36 378-379 comparison with rubredoxin, 36 404 coordinated amino acid spacing, 36 399 cucumber basic protein, 36 390 electron transfer routes, 36 403-404 electron transport, 36 378 EXAFS studies, 36 390-391 functional role, 36 382-383 occurrence, 36 379-382 properties, 36 380 pseudoazurin, 36 389-390 reduction potentials, 36 393-396 self-exchange rate constants, 36 401-403 UV-VIS spectra, 36 391-393 Blue species... 

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