Structural and Functional Characterization of IclR Transcription Regulators
This work is a part of a large project in our laboratory that is aimed toward characterization of prokaryotic transcription regulators from different families and their interactions with small-molecule effectors. My study was focused of IclR family of transcriprion regulators, specifically on its fo...
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ndltd-LACETR-oai-collectionscanada.gc.ca-OTU.1807-182912013-11-02T03:43:30ZStructural and Functional Characterization of IclR Transcription RegulatorsEzersky, AlexandraTranscription regulatorsX-ray crystallography0760This work is a part of a large project in our laboratory that is aimed toward characterization of prokaryotic transcription regulators from different families and their interactions with small-molecule effectors. My study was focused of IclR family of transcriprion regulators, specifically on its founding member Isocytrate Lyase Regulator (IclR) from E.coli and AllR regulator from E.coli, which share 42% sequence identity with IclR. I used a combination of biophysical, biochemical and structural biology techniques to explore the mechanisms by which IclR and AllR interact with their effectors. I performed site-directed mutagenesis experiments in order to research the role of individual amino acids in interaction of AllR regulator with its previously identified effector glyoxylate and to test whether oligomerization plays a role in effector-induced signal transduction by AllR. Using differential light scattering, which allows high-throughput screening of small molecules for thermostabilization of proteins, I identified potential effctors for the IclR regulator. The physiological relevance of these candidate molecules was tested in-vitro and in-vivo and their interaction with IclR was characterized by Isothermal Titration Calorimetry and X-ray Crystallography.Edwards, Aled M.2009-112010-01-15T19:17:22ZNO_RESTRICTION2010-01-15T19:17:22Z2010-01-15T19:17:22ZThesishttp://hdl.handle.net/1807/18291en_ca |
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Transcription regulators X-ray crystallography 0760 |
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Transcription regulators X-ray crystallography 0760 Ezersky, Alexandra Structural and Functional Characterization of IclR Transcription Regulators |
description |
This work is a part of a large project in our laboratory that is aimed toward characterization of prokaryotic transcription regulators from different families and their interactions with small-molecule effectors. My study was focused of IclR family of transcriprion regulators, specifically on its founding member Isocytrate Lyase Regulator (IclR) from E.coli and AllR regulator from E.coli, which share 42% sequence identity with IclR. I used a combination of biophysical, biochemical and structural biology techniques to explore the mechanisms by which IclR and AllR interact with their effectors.
I performed site-directed mutagenesis experiments in order to research the role of individual amino acids in interaction of AllR regulator with its previously identified effector glyoxylate and to test whether oligomerization plays a role in effector-induced signal transduction by AllR. Using differential light scattering, which allows high-throughput screening of small molecules for thermostabilization of proteins, I identified potential effctors for the IclR regulator. The physiological relevance of these candidate molecules was tested in-vitro and in-vivo and their interaction with IclR was characterized by Isothermal Titration Calorimetry and X-ray Crystallography. |
author2 |
Edwards, Aled M. |
author_facet |
Edwards, Aled M. Ezersky, Alexandra |
author |
Ezersky, Alexandra |
author_sort |
Ezersky, Alexandra |
title |
Structural and Functional Characterization of IclR Transcription Regulators |
title_short |
Structural and Functional Characterization of IclR Transcription Regulators |
title_full |
Structural and Functional Characterization of IclR Transcription Regulators |
title_fullStr |
Structural and Functional Characterization of IclR Transcription Regulators |
title_full_unstemmed |
Structural and Functional Characterization of IclR Transcription Regulators |
title_sort |
structural and functional characterization of iclr transcription regulators |
publishDate |
2009 |
url |
http://hdl.handle.net/1807/18291 |
work_keys_str_mv |
AT ezerskyalexandra structuralandfunctionalcharacterizationoficlrtranscriptionregulators |
_version_ |
1716612292293951488 |