Purification and Characterization of Novel Denitrosylases from Yeast and Mammals
<p>S-nitrosylation, the prototypic mechanism of redox-based signal transduction, involves the covalent attachment of a nitrogen monoxide group to a Cys-thiol side chain. S-nitrosylation of proteins has been demonstrated to affect a broad range of functional parameters including enzymatic activ...
Main Author: | |
---|---|
Other Authors: | |
Published: |
2012
|
Subjects: | |
Online Access: | http://hdl.handle.net/10161/6165 |
id |
ndltd-DUKE-oai-dukespace.lib.duke.edu-10161-6165 |
---|---|
record_format |
oai_dc |
spelling |
ndltd-DUKE-oai-dukespace.lib.duke.edu-10161-61652016-01-07T03:29:54ZPurification and Characterization of Novel Denitrosylases from Yeast and MammalsAnand, PuneetBiochemistryDenitrosylasesNitric OxideNitric oxide synthasesS-nitrosylation<p>S-nitrosylation, the prototypic mechanism of redox-based signal transduction, involves the covalent attachment of a nitrogen monoxide group to a Cys-thiol side chain. S-nitrosylation of proteins has been demonstrated to affect a broad range of functional parameters including enzymatic activity, subcellular localization, protein-protein interactions and protein stability. The primary focus of my dissertation was to solve a problem of great importance in the field of S-nitrosylation, which is, to identify denitrosylase(s) i.e., enzymes that remove NO groups from S-nitrosothiols. Recent progress in elucidating the cellular regulation of S-nitrosylation has led to the identification of two physiologically relevant denitrosylating activities that remove the NO group from S-nitrosylated substrates. Thioredoxin/thioredoxin reductase (Trx system) functions as an NADPH-dependent denitrosylase across a broad range of S-nitrosylated proteins (SNO-proteins). S-nitroso-glutathione reductase (GSNOR), which is highly conserved across phylogeny, metabolizes GSNO utilizing NADH as a reducing coenzyme, thereby shifting equilibria between GSNO and SNO-proteins. This dissertation describes the discovery of two novel denitrosylases: one from yeast and the other from mammals. Using technique of column chromatography we have purified these novel denitrosylases to homogeneity and have demonstrated a principal contribution of these enzymes towards S-nitrosothiol metabolism.</p>DissertationStamler, Jonathan SKreuzer, Kenneth N2012Dissertationhttp://hdl.handle.net/10161/6165 |
collection |
NDLTD |
sources |
NDLTD |
topic |
Biochemistry Denitrosylases Nitric Oxide Nitric oxide synthases S-nitrosylation |
spellingShingle |
Biochemistry Denitrosylases Nitric Oxide Nitric oxide synthases S-nitrosylation Anand, Puneet Purification and Characterization of Novel Denitrosylases from Yeast and Mammals |
description |
<p>S-nitrosylation, the prototypic mechanism of redox-based signal transduction, involves the covalent attachment of a nitrogen monoxide group to a Cys-thiol side chain. S-nitrosylation of proteins has been demonstrated to affect a broad range of functional parameters including enzymatic activity, subcellular localization, protein-protein interactions and protein stability. The primary focus of my dissertation was to solve a problem of great importance in the field of S-nitrosylation, which is, to identify denitrosylase(s) i.e., enzymes that remove NO groups from S-nitrosothiols. Recent progress in elucidating the cellular regulation of S-nitrosylation has led to the identification of two physiologically relevant denitrosylating activities that remove the NO group from S-nitrosylated substrates. Thioredoxin/thioredoxin reductase (Trx system) functions as an NADPH-dependent denitrosylase across a broad range of S-nitrosylated proteins (SNO-proteins). S-nitroso-glutathione reductase (GSNOR), which is highly conserved across phylogeny, metabolizes GSNO utilizing NADH as a reducing coenzyme, thereby shifting equilibria between GSNO and SNO-proteins. This dissertation describes the discovery of two novel denitrosylases: one from yeast and the other from mammals. Using technique of column chromatography we have purified these novel denitrosylases to homogeneity and have demonstrated a principal contribution of these enzymes towards S-nitrosothiol metabolism.</p> === Dissertation |
author2 |
Stamler, Jonathan S |
author_facet |
Stamler, Jonathan S Anand, Puneet |
author |
Anand, Puneet |
author_sort |
Anand, Puneet |
title |
Purification and Characterization of Novel Denitrosylases from Yeast and Mammals |
title_short |
Purification and Characterization of Novel Denitrosylases from Yeast and Mammals |
title_full |
Purification and Characterization of Novel Denitrosylases from Yeast and Mammals |
title_fullStr |
Purification and Characterization of Novel Denitrosylases from Yeast and Mammals |
title_full_unstemmed |
Purification and Characterization of Novel Denitrosylases from Yeast and Mammals |
title_sort |
purification and characterization of novel denitrosylases from yeast and mammals |
publishDate |
2012 |
url |
http://hdl.handle.net/10161/6165 |
work_keys_str_mv |
AT anandpuneet purificationandcharacterizationofnoveldenitrosylasesfromyeastandmammals |
_version_ |
1718160427579342848 |