Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.612561
Title: Synthesis and evaluation of stable analogues of [tau]-Phosphohistidine
Author: McAllister , Tom
Awarding Body: University of Leeds
Current Institution: University of Leeds
Date of Award: 2012
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Abstract:
t-Phosphohistidine is an acid-labile post-translational modification found in some proteins. Study of this modification has been impaired by a lack of biochemical tools to identify proteins contai ning t-phosphohistidine and ways to study protein-prolein interactions mediated by 't-phosphohistidinc. A t -phosphohistidine analogue based upon a triazole scaffold has been synthesised. which replicates the oricnlal ion of functional groups in t -phosphohistidine and this has been shown to mimic l-phosphohistidinc in biological systems. This was achieved through the synthesis of peptides containing I which required production of suitably protected building block. It was found that following incorporation into peptide, the ethyl protecting groups (R= El) initially used on the phosphonate were not removed by trinuoroacetic acid-based cleavage conditions and required a subsequent deprotection step. A ncw synthetic roule to the precursor alkynylphosphonate was developed to allow incorporati on of benzyl and tert-butyl phosphonate protecting groups (R= Bnr IB u) which allowed peptides containing analogue I to be synthesised directly.Peptides synthesised by this method have been used to recapitulate the binding of a histidine phosphorylated protein, phosphoenolpyruvate synthetase, with its cognate regulatory protein YdiA and reveal the promiscuity of growth factor receptor-bound protein 2 (a phosphotyrosine-binding protein) to bind t-phosphohistidine. Peptides containing I have also been used to develop an immunochemical tool (through development of an antibody) that could be used to detect -r-phosphohistidine and in a selection screen to identify a peptide aptamer that binds to the analogue; a non-antibody phosphothreonine-binding protein was identified in this process.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.612561  DOI: Not available
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