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Purification and Characterization of Novel Denitrosylases from Yeast and Mammals

dc.contributor.advisor Stamler, Jonathan S
dc.contributor.advisor Kreuzer, Kenneth N
dc.contributor.author Anand, Puneet
dc.date.accessioned 2013-01-16T20:29:04Z
dc.date.available 2016-01-06T05:30:04Z
dc.date.issued 2012
dc.identifier.uri https://hdl.handle.net/10161/6165
dc.description.abstract <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>
dc.subject Biochemistry
dc.subject Denitrosylases
dc.subject Nitric Oxide
dc.subject Nitric oxide synthases
dc.subject S-nitrosylation
dc.title Purification and Characterization of Novel Denitrosylases from Yeast and Mammals
dc.type Dissertation
dc.department Biochemistry
duke.embargo.months 36


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