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"Development of selective antagonists and subtype .. (SELECTIPROBE)
"Development of selective antagonists and subtype selective ligands for the D-myo-inositol 1,4,5-trisphosphate receptors."
(SELECTIPROBE)
Date du début: 1 juin 2012,
Date de fin: 31 mai 2014
PROJET
TERMINÉ
"The overall aim of this project is the development of novel ligands to probe the role of D-myo-inositol 1,4,5-trisphosphate receptors (InsP3Rs) in intracellular calcium signalling. Two key goals are the development of selective and potent non-inositol InsP3R antagonists and the development of ligands that bind selectively to one of the three subtype of InsP3Rs. Using hits from an in silico screen, we will develop and then employ thermal shift analysis, surface plasmon resonance and isothermal titration calorimetry assays to identify promising lead compounds. We will determine whether these compounds act as agonists or antagonists using a fluorescence-based sea urchin egg homogenate assay before data are obtained in further, more intact, cellular assays. In order to facilitate the use of these compounds in cells they will be rendered cell permeant using previously developed technology. X-ray crystallographic studies with these compounds, in particular the antagonists, will be undertaken in order to shed light the molecular mechanism of InsP3R activation. Subtype selective ligands will be useful in determining the particular role of each of the InsP3R subtypes in normal cellular function and cellular dysfunction, which underlies diseases such as heart arrhythmia and cardiac hypertrophy. This project has been designed to capitalise on the skills of the fellow in chemical synthesis and her knowledge of phosphate bioisosteres. Further advanced training will be provided in chemical synthesis, biophysical techniques and molecular biology in addition to cooperation with industrial collaborators Inhibox. Consequently, the fellow will significantly enhance and diversify her skills in this highly multidisciplinary project."
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