Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.750480
Title: Studies of the quinone binding sites of the Escherichia coli terminal oxidases, cytochromes bo3 & bd
Author: Hastings, Stuart Fairbairn
Awarding Body: University of St Andrews
Current Institution: University of St Andrews
Date of Award: 1997
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Abstract:
The structure and function relationships involved in the binding of quinones to the terminal oxidases of Escherichia coli, cytochromes bd and bo3, were investigated using redox potentiometery, site-directed mutagenesis and magnetic resonance techniques. A stable semiquinone was identified as an intermediate of quinol oxidation by cytochrome bd in appropriately poised samples of both the membrane-bound and purified enzyme reconstituted with excess ubi- and menaquinone analogues. The effects of the inhibitors HOQNO and aurachin D on semiquinone formation were assessed. The potentiometric behaviour of the semiquinone stabilised by membrane-bound and purified cytochrome bo3, reconstituted with excess quinone analogues was characterised. The effects of two cytochrome bo3 subunit II mutations and the inhibitor tridecylstigmatellin were studied. The data presented is consistent with the presence of one quinone binding site. The hyperfine splittings present in the ESR spectrum of the cytochrome bo3 semiquinone were resolved by ENDOR spectroscopy. The resultant electronic structure of the bound semiquinone and the nature of the hydrogen bonding to the protein is described. ESEEM spectroscopy was used to identify a nitrogen nucleus hydrogen bonded to the semiquinone. A model of this quinone binding site and a possible location is presented.
Supervisor: Ingledew, John Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.750480  DOI: Not available
Keywords: QP671.C85H2 ; Cytochromes
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