Selection of cyclic peptide inhibitors of hypoxia-inducible factor
Selection of cyclic peptide inhibitors of hypoxia-inducible factor
Hypoxia-inducible factor (HIF) is a heterodimer that is formed from the association of HIF-? and HIF-1? subunits. HIF acts as a potent inducer of genes involved in angiogenesis and neovascularisation. Three different variants of HIF have been identified to date; HIF-1 and HIF-2 are thought to play key roles in tumour blood vessel formation while with HIF-3 is thought to act as a negative regulator of the hypoxic response.
In this study a genetic selection methodology is employed that combines a bacterial reverse two-hybrid system with SICLOPPS (split-intein circular ligation of peptides and proteins), a protocol for the construction of libraries of around a hundred million cyclic peptides using split-inteins in vivo. After construction of the appropriate reverse twohybrid systems, several SICLOPPS libraries were screened for inhibitors of HIF-1 and HIF-2 heterodimerisation. After a number of rounds of secondary screening, the activity of the remaining compounds was ranked by using the HIF reverse two-hybrid systems. The most potent compounds were synthesised as Tat-tagged derivatives for further characterisation in a luciferase based mammalian cell assay. The most potent cyclic peptide exhibited IC50s ranging from 26 to 106 ?M in U2OS and MCF7 cell lines.
Nordgren, Ida Karin
3ccc4429-3f5a-4b52-8d56-b24220b45bba
29 December 2011
Nordgren, Ida Karin
3ccc4429-3f5a-4b52-8d56-b24220b45bba
Tavassoli, Ali
d561cf8f-2669-46b5-b6e1-2016c85d63b2
Nordgren, Ida Karin
(2011)
Selection of cyclic peptide inhibitors of hypoxia-inducible factor.
University of Southampton, School of Chemistry, Doctoral Thesis, 295pp.
Record type:
Thesis
(Doctoral)
Abstract
Hypoxia-inducible factor (HIF) is a heterodimer that is formed from the association of HIF-? and HIF-1? subunits. HIF acts as a potent inducer of genes involved in angiogenesis and neovascularisation. Three different variants of HIF have been identified to date; HIF-1 and HIF-2 are thought to play key roles in tumour blood vessel formation while with HIF-3 is thought to act as a negative regulator of the hypoxic response.
In this study a genetic selection methodology is employed that combines a bacterial reverse two-hybrid system with SICLOPPS (split-intein circular ligation of peptides and proteins), a protocol for the construction of libraries of around a hundred million cyclic peptides using split-inteins in vivo. After construction of the appropriate reverse twohybrid systems, several SICLOPPS libraries were screened for inhibitors of HIF-1 and HIF-2 heterodimerisation. After a number of rounds of secondary screening, the activity of the remaining compounds was ranked by using the HIF reverse two-hybrid systems. The most potent compounds were synthesised as Tat-tagged derivatives for further characterisation in a luciferase based mammalian cell assay. The most potent cyclic peptide exhibited IC50s ranging from 26 to 106 ?M in U2OS and MCF7 cell lines.
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Published date: 29 December 2011
Organisations:
University of Southampton, Chemistry
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Local EPrints ID: 344931
URI: http://eprints.soton.ac.uk/id/eprint/344931
PURE UUID: 9ff6d476-0b7d-4029-9d42-e8bcb0b8bdef
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Date deposited: 25 Feb 2013 14:29
Last modified: 15 Mar 2024 03:26
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Author:
Ida Karin Nordgren
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