Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.663113
Title: Control mechanisms of intra-molecular electron transfer in oxidoreductases
Author: Turner, Karen L.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1999
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Abstract:
The intra-molecular electron transfer steps in three multi-centred oxidoreductases were studied by redox potentiometry and protein film voltammetry. Cytochrome cd1 is a disimmilatory nitrite reductase, the achieves the reduction of nitrite to nitric oxide. Reduction of the enzyme from Thiosphaera pantotropha is associated with a conformational change (Williams et al., 1997) and hysteresis is observed in the potentiometric titration at room temperature (Kobayashi et al., 1997). The potentials in the oxidative and reductive directions are separated by 150 mV at 20°C but ?DE decreases at high temperatures. The increased reversibility at higher temperatures is also shown by a rise in the n-value and the appropriate spectral response when the potential is perturbed in both the oxidative and reductive directions. At the highest temperatures, when equilibrium conditions are reached, the interconversion of the completely oxidised and completely reduced states is characterised by a potential of 140 mV and an n-value of 2. At 20 °C the electron transfer is conformationally gated such that a proposed square scheme is essentially a one-way system. As the temperature is raised, the rate of the conformational change increases and the interconversion approaches reversibility within the measurement time. Flavocytochrome P-450 BM3 is a fatty-acid monooxygenase from Bacillus megaterium. The complete redox characterisation of the enzyme has been achieved by potentiometric titrations on the intact holoenzyme and its component domains. The reduction potential of the haem is increased by more than 100 mV on fatty-acid substrate binding at the active site (Em = -368 ± 6 mV, substrate-free, to Em = -239 ± 6 mV, arachidonate-bound). The FMN to haem electron-transfer is under thermodynamic control, such that the futile cycling of electrons and hydrogen peroxide formation is prevented. The blue FAD semiquinone species is observed during the course of the reductive titrations of these enzymes (E1 = -283 mV).
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.663113  DOI: Not available
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