Strategies for characterisation of the unexplored human phosphoproteome

Hardman, GE (2017) Strategies for characterisation of the unexplored human phosphoproteome. PhD thesis, University of Liverpool.

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Abstract

Post-translational modification of proteins by addition of a phosphate group serves as a means of rapidly regulating protein function, and thus has vital roles in all aspects of cell biology. Phosphorylation of serine, threonine and tyrosine are the focus of the vast majority of studies aimed at elucidating the function of such modification in human cells. However, there is growing evidence that phosphohistidine (pHis) may also have a role in diverse cellular processes in mammalian systems. Characterisation of pHis is compromised by the inherent instability of the acid-labile phosphoramidate bond, meaning there is a severe lack of suitable experimental strategies to pinpoint the presence of pHis in proteins. The acidic conditions integral to current phosphoproteomics workflows are incompatible with analysis of acid-labile pHis. In this work, chemical phosphorylation was used to generate a set of model pHis-containing peptides, for which stability of the histidine phosphorylation across a range of pH values and at elevated temperature was assessed. The pHis-containing peptides were further used to evaluate various existing enrichment strategies for their suitability for pHis. None of the enrichment methods ... (continues)

Item Type:	Thesis (PhD)
Divisions:	Faculty of Health and Life Sciences > Faculty of Health and Life Sciences
Depositing User:	Symplectic Admin
Date Deposited:	16 Aug 2018 13:43
Last Modified:	17 Aug 2024 22:25
DOI:	10.17638/03018266
Supervisors:	Eyers, Claire
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3018266