Reflection anisotropy spectroscopy as a potential new tool for linking macromolecular conformation to biological function

Convery, James
Reflection anisotropy spectroscopy as a potential new tool for linking macromolecular conformation to biological function. Unspecified thesis, University of Liverpool.

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The results reported in this thesis were obtained in a research programme that employed the surface sensitive optical probe reflection anisotropy spectroscopy (RAS) and related techniques to investigate conformational change associated with electron transfer processes in the protein cytochrome P450 reductase (CPR) adsorbed onto the Au(110)/electrode interface. It was found that controlling the adsorption of an ordered layer of CPR on the Au(110) surface is difficult and far more complex than first expected. In order to understand the adsorption process further an extensive investigation using quartz crystal microbalance with dissipation (QCM D) was carried out. The results of the QCM D experiments established that the conditions necessary for the adsorption of a monolayer of CPR are extremely sensitive to protein concentration and buffer pH. This was a crucial step in the research which allowed the preparation of adsorbed ordered monolayers of CPR on Au(110)/electrode interfaces. The RA spectral signatures of a monolayer and bilayer of CPR adsorbed on the Au(110) surface have been identified and are reported in this thesis. Using azimuthal dependent reflection anisotropy spectroscopy (ADRAS) it was shown that a monolayer of adsorbed CPR aligns along one of the principle axis of the Au(110) surface. RAS investigations of the effect of inducing electron transfer processes in an adsorbed monolayer of CPR by stepping the applied electrode potential reveal changes in RA spectral shape and intensity which may be associated with conformational events in the adsorbed CPR. The RA spectra produced as a function of applied potential show that the adsorption of a monolayer of CPR impedes the Au(110) surface reconstruction from a (13) to the anion induced (11) structures.

Item Type: Thesis (Unspecified)
Additional Information: Date: 2012-08 (completed)
Subjects: ?? QC ??
Divisions: Faculty of Science and Engineering > School of Physical Sciences
Depositing User: Symplectic Admin
Date Deposited: 07 Aug 2013 10:08
Last Modified: 17 Dec 2022 01:32
DOI: 10.17638/00009473