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Title: Analysis of the proteomic effects of leukaemic protein tyrosine kinases
Author: Lee, Chia-Fang
ISNI:       0000 0001 3606 9059
Awarding Body: University of Manchester
Current Institution: University of Manchester
Date of Award: 2008
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There are a number of leukaemogenic protein tyrosine kinases (PTKs) associated with leukaemic transfonnation. Whilst each is linked with a specific disease their functional activity poses the question whether they have a degree of commonality in their effects upon target cells. Exon array analysis of the effects of 6 leukaemogenic PTKs (BCRJABL, TELIPDGFRp, FIPIIPDGFRa, D816V KIT, NPMlALK and FLT3ITD) revealed few common effects on the transcriptome. It is apparent, however, that proteome changes are not directly governed by transcriptome changes. Therefore, a new generation of eight channel iTRAQ reagent was applied to analyse the effects of these 6 leukaemogenic PTKs. Again these were found to have disparate effects on the proteome with few common targets. BCRJABL had the greatest effect on the proteome and had more effects in common with FIPIIPDGFRa. The only protein commonly affected by all six oncogenes was eosinophil-associated ribonuclease 7. Furthennore, correlation of the proteomics data with that from the exon microarray not only showed poor levels of correlation between transcript changes and protein level changes, but also revealed alternative patterns of regulation of the CAPG protein by different oncogenes, illustrating the utility of such a combined approach. The study was then focused on investigating the changes within the phosphoproteome. Due to the low stoichiometry of phosphorylation, some level of enrichment IS required. A method using phosphoprotein enrichment in conjunction with iTRAQ based mass spectrometry was developed using BalF3 and BCRJABL BalF3 cell lines. Following data validation, 27 significantly changing proteins were identified. The results revealed that more than half of the proteins showed statistically significant differences in phosphoprotein enrichment fractions remain unchanged in protein expression levels in whole cell lysates, including eIF4E. eIF4E has been found its phosphorylation status is critical in tumourgenesis. The data suggest this approach will be useful for the study of phosphorylation events.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available