Redox thermodynamic data provide a detailed insight into control of the reduction potential E-o' of the [Fe(S-Cys)(4)] site in rubredoxin. Mutant forms were studied in which specific structural changes were made in both the primary and secondary coordination spheres. Those changes have been probed by resonance Raman spectroscopy. The decrease of similar to 200mV in E-o' observed for the [Fe(S-Cys)(3)(O-Ser)](-/2-) couples in the surface ligand mutants C9S and S42S is essentially enthalpic in origin and associated with the substitution of ligand thiolate by ligand elate. However, the pH dependence of the potentials below characteristic pK(a)(red)approximate to7 is an entropic contribution, plausibly associated with increased conformational flexibility induced by a longer Fe-II-O(H)-Ser bond in the reduced form. The presence of a second surface Ser ligand in the new double mutant protein C9S/C42S affects the enthalpic term primarily for pH>pK(a)(red)greater than or equal to9.3, but for pH<pK(a)(red) the entropic term again becomes significant. The available data for the internal ligand mutant C39S appear to follow those for the surface ligand mutants. A longer Fe-III-O-Ser link in the oxidized form is expected from structural considerations and a smaller decrease of similar to 100 mV in E-o' is observed for this particular [Fe(S-Cys)(3)(O-Ser)](-/2-) couple. The reduced form is particularly susceptible to hydrolysis with consequent irreversible electrochemistry, an apparent consequence of an even longer Fe-II-O(H)-Ser bond. The second internal ligand mutant C6S exhibits starkly different behaviour. The cause was traced to the presence of a Fe-III(SCys)(3)(0H)(-) centre in the oxidized form. Resonance Raman spectroscopy on Fe-54-, (H2O)-O-18- and D2O-enriched samples supports the presence of an iron hydroxyl group that donates a hydrogen bond to the OH group of the free S6 side chain and/or a cluster of bound water molecules. The thermodynamic parameters can then be rationalized in terms of the following processes: pH>pK(a)similar to9: [Fe-III(S-Cys)(3)(OH)](-) + e(-) --> [Fe-II(S-Cys)(3)(OH>)](2-) pH<pK(a): [Fe-III(S-Cys)(3)(OH)](-) + H+ + e(-) <right arrow> [Fe-II(S-Cys)(3)(OH2)](-).
Attenzione! Scheda prodotto non ancora validata dall'Ateneo
Dati e metadati della pubblicazione sono in fase di verifica da parte dell'Ateneo. In caso di errori o violazione dei diritti d'autore, contattare: firstname.lastname@example.org
|Anno di pubblicazione:||2001|
|Titolo:||Redox thermodynamics of mutant forms of the rubredoxin from Clostridiumpasteurianum: identification of a stable Fe-III(S-Cys)(3)(OH) centre in the C6S mutant|
|Autori:||Xiao ZG; Gardner AR; Cross M; Maes EM; Czernuszewicz RS; Sola M; Wedd AG|
|Autori interni:||SOLA, Marco|
|Appare nelle tipologie:||Articolo su rivista|
File in questo prodotto:
I documenti presenti in Iris Unimore sono rilasciati con licenza Creative Commons Attribuzione - Non commerciale - Non opere derivate 3.0 Italia, salvo diversa indicazione.
In caso di violazione di copyright, contattare Supporto Iris