Fragment screening for stabilizers and binders to human DJ-1

Parkinson’s disease (PD) is a chronic progressive neurodegenerative disease that is on the rise. Currently, treatments are only available to alleviate the span of symptoms but not to cure the morbidity. This project focuses on finding ligands to increase the stability of DJ1 where its anti-oxidant p...

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Main Author: Oh, Xu Xuan
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Format: Final Year Project
Language:English
Published: Nanyang Technological University 2016
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Online Access:http://hdl.handle.net/10356/68921
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Institution: Nanyang Technological University
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spelling sg-ntu-dr.10356-689212023-02-28T18:03:31Z Fragment screening for stabilizers and binders to human DJ-1 Oh, Xu Xuan - School of Biological Sciences Susana Geifman Shochat SGeifman@ntu.edu.sg DRNTU::Science::Biological sciences::Human anatomy and physiology::Neurobiology DRNTU::Science::Biological sciences::Biophysics Parkinson’s disease (PD) is a chronic progressive neurodegenerative disease that is on the rise. Currently, treatments are only available to alleviate the span of symptoms but not to cure the morbidity. This project focuses on finding ligands to increase the stability of DJ1 where its anti-oxidant properties help to reduce the rate of dopaminergic neurons degeneration, delaying onset of PD. The binding of fragments to DJ1 constructs are screened using biophysical techniques. ThermoFluor assay (DSF) is used to screen the fragment libraries where five ligands namely CPD1, CPD2, CPD3, CPD4 and CPD5 were identified to confer stability to DJ1. The assays were performed under control conditions and oxidative conditions from 0x to 20x hydrogen peroxide (H2O2) molarity in excess. In the presence of CPD1, DJ1 was observed to have a positive melting temperature (ΔTm) shift in oxidised DJ1, showing an enhancement of DJ1 stability under oxidative conditions. Isothermal titration calorimetry (ITC) conducted between DJ1 and CPD5 shows evidence of high binding affinity with DJ1. Experiments performed with CPD1 run need to be validated. To learn about the binding sites for the fragments in DJ1, hanging-drop crystallization is attempted and suitable cultured crystals will undergo X-ray crystallography analysis. Bachelor of Science in Biomedical Sciences 2016-08-04T02:58:28Z 2016-08-04T02:58:28Z 2016 Final Year Project (FYP) http://hdl.handle.net/10356/68921 en 30 p. application/pdf Nanyang Technological University
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science::Biological sciences::Human anatomy and physiology::Neurobiology
DRNTU::Science::Biological sciences::Biophysics
spellingShingle DRNTU::Science::Biological sciences::Human anatomy and physiology::Neurobiology
DRNTU::Science::Biological sciences::Biophysics
Oh, Xu Xuan
Fragment screening for stabilizers and binders to human DJ-1
description Parkinson’s disease (PD) is a chronic progressive neurodegenerative disease that is on the rise. Currently, treatments are only available to alleviate the span of symptoms but not to cure the morbidity. This project focuses on finding ligands to increase the stability of DJ1 where its anti-oxidant properties help to reduce the rate of dopaminergic neurons degeneration, delaying onset of PD. The binding of fragments to DJ1 constructs are screened using biophysical techniques. ThermoFluor assay (DSF) is used to screen the fragment libraries where five ligands namely CPD1, CPD2, CPD3, CPD4 and CPD5 were identified to confer stability to DJ1. The assays were performed under control conditions and oxidative conditions from 0x to 20x hydrogen peroxide (H2O2) molarity in excess. In the presence of CPD1, DJ1 was observed to have a positive melting temperature (ΔTm) shift in oxidised DJ1, showing an enhancement of DJ1 stability under oxidative conditions. Isothermal titration calorimetry (ITC) conducted between DJ1 and CPD5 shows evidence of high binding affinity with DJ1. Experiments performed with CPD1 run need to be validated. To learn about the binding sites for the fragments in DJ1, hanging-drop crystallization is attempted and suitable cultured crystals will undergo X-ray crystallography analysis.
author2 -
author_facet -
Oh, Xu Xuan
format Final Year Project
author Oh, Xu Xuan
author_sort Oh, Xu Xuan
title Fragment screening for stabilizers and binders to human DJ-1
title_short Fragment screening for stabilizers and binders to human DJ-1
title_full Fragment screening for stabilizers and binders to human DJ-1
title_fullStr Fragment screening for stabilizers and binders to human DJ-1
title_full_unstemmed Fragment screening for stabilizers and binders to human DJ-1
title_sort fragment screening for stabilizers and binders to human dj-1
publisher Nanyang Technological University
publishDate 2016
url http://hdl.handle.net/10356/68921
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