Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.582851
Title: Development of an enrichment procedure for the isolation and analysis of nuclear proteins
Author: Foulger , Larus E.
Awarding Body: Liverpool John Moores University
Current Institution: Liverpool John Moores University
Date of Award: 2012
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Abstract:
The presence of some nuclear proteins (HMGBs included) has been found to be associated with a variety of diseases including several cancers and inflammatory disorders, including sepsis. The presence of these proteins, and their associated PTMs, are the source of a large amount of research to identify the specific modifications that make these chromatin regulating proteins the source of terminal disorders. The initial step was the determination of lysis conditions that created the optimal conditions for the separation of nuclear proteins from the chromatin structure in 'mild' conditions, using KCl/phosphate, in their native state. The use of these buffers and slight variation in their concentrations and ratios allowed for an enrichment of soluble proteins and the precipitation of non-soluble ones. This would, therefore, allow for the isolation of native proteins, and any protein-protein interactions that were present. This then allowed for the application of this method to nuclei and the extraction of proteins in their native state whose interactions with proteins that had eo-eluted via ion-exchange. Allowing for the identification and isolation of several possible protein-protein interactions associated with chromatin proteins. The use of cross-linking experiments allowed for the determination of non-transient interactions between one of these complexes, showing an interaction between HMGB 1 and Cyclophilin B, a peptide prolyl isomerase, most often found in the Endoplasmic Reticulum of the cytoplasm. The application of this enrichment method to chicken blood- plasma allowed for the precipitation of significant amounts of 'minimally- soluble' proteins (including immunonglubulins and fibrinogens), whilst leaving the most soluble ones present (including apolipoprotein).
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.582851  DOI: Not available
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