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Title: The application of mass spectrometric analysis to biochemical studies of the role and function of cyclic CMP
Author: Bond, A. E.
Awarding Body: University of Wales Swansea
Current Institution: Swansea University
Date of Award: 2006
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This study investigates the identities of modified nucleosides from cancer patients’ urine. This was conducted using a Q – ToF MS and accurate mass program to determine the fragmentation pathway of each nucleoside investigated. The identities of the modified nucleosides confirmed in this study are: methyl-acetyl cytidine, 5’-O-methyl cytidine, ethyl-acetyl cytidine and 5’ – formylcytidine. The fragmentation profile of pseudouridine is also investigated as this nucleoside has been found in various types of cancer. With several of the elevated modified nucleosides in cancer patients’ urine being cytidine compounds, and given the elevation of cCMP levels in hyperproliferating cells, this study also considers the fragmentation pathways of cCMP analogous by using the fragmentation pathways of chlorinated adenine compounds: 8-Cl –adenine, - adenosine, - 5’AMP and Sp/Rp-8-Cl-cAMPS. Q-ToF MS was utilised for accurate mass analysis and LCQ- Deca ESI MS for MSn analysis. A proposed theoretical fragmentation pathway has been designed for 5-Cl-3’,5’-cCMP from the known pathways and investigated profiles in this study. The identification of the proteins specifically phosphorylated in response to cCMP is also investigated in this study. Murine brain tissue was utilised and following incubation with cyclic nucleotides: cyclic AMP, cyclic GMP, cyclic CMP and control (blank), these preparations were initially analysed using 2-D gel electrophoresis. However, there was insufficient acquisition of data to generate any conclusive deductions, so the continuation of the studies took place with the use of other techniques i.e. immobilised metal (ion) affinity chromatography and mass spectrometry.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available