ERA

Download the full-sized PDF of Protein tyrosine nitration in mast cellsDownload the full-sized PDF

Analytics

Share

Permanent link (DOI): https://doi.org/10.7939/R3FS63

Download

Export to: EndNote  |  Zotero  |  Mendeley

Communities

This file is in the following communities:

Graduate Studies and Research, Faculty of

Collections

This file is in the following collections:

Theses and Dissertations

Protein tyrosine nitration in mast cells Open Access

Descriptions

Other title
Subject/Keyword
mast cells
nitric oxide
protein tyrosine nitration
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Sekar, Yokananth
Supervisor and department
Befus, A. Dean (Medicine)
Examining committee member and department
Olson, David M. (Physiology)
Davidge, Sandra T. (Obstetrics & Gynecology and Physiology)
Cameron, Lisa (Medicine)
Schulz, Richard (Pediatrics and Pharmacology)
Kubes, Paul (Physiology & Pharmacology and Medicine, University of Calgary)
Department
Medicine
Specialization

Date accepted
2011-01-11T17:36:33Z
Graduation date
2011-06
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
Nitric oxide (NO) is a short-lived free radical that plays a critical role in the regulation of cellular signalling. Mast cell (MC) derived NO and exogenous NO regulate MC activities including the inhibition of MC degranulation. At a molecular level the intermediate metabolites of NO modify protein structure and function through several mechanisms including protein tyrosine nitration. To begin to elucidate the molecular mechanisms underlying the effects of NO in MC, we investigated protein tyrosine nitration in human mast cell lines HMC-1 and LAD2 treated with the NO donor S-nitrosoglutathione (SNOG). Using two dimensional gel western blot analysis with an anti-nitrotyrosine antibody together with mass spectroscopy we identified aldolase A, an enzyme of the glycolytic pathway, as a target for tyrosine nitration in MC. S-nitrosoglutathione treatment also reduced the Vmax of aldolase in HMC-1 and LAD2. Nuclear magnetic resonance (NMR) analysis showed that despite these changes in activity of a critical enzyme in glycolysis, there was no significant change in total cellular ATP content, although the AMP/ATP ratio was altered. Elevated levels of lactate and pyruvate suggested that SNOG treatment enhanced glycolysis. Reduced aldolase activity was associated with increased intracellular levels of its substrate, fructose-1,6-bisphosphate (FBP). Interestingly, FBP inhibited IgE-mediated MC degranulation and intracellular Ca2+ levels in LAD2 cells. In addition to aldolase, 15-hydroxy prostaglandin dehydrogenase (PGDH), a critical enzyme in the metabolism of PGE2, was identified as a prominent target for tyrosine nitration in LAD2 cells. Thus for the first time we report evidence of protein tyrosine nitration in human MC lines and identify aldolase A as a prominent target in HMC-1 and LAD2; and PGDH in LAD2 cells. The post translational nitration of aldolase A and PGDH may be important pathways that regulate MC phenotype and function.
Language
English
DOI
doi:10.7939/R3FS63
Rights
License granted by Yokananth Sekar (yokananth@ualberta.ca) on 2011-01-09T23:22:33Z (GMT): Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of the above terms. The author reserves all other publication and other rights in association with the copyright in the thesis, and except as herein provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
Citation for previous publication

File Details

Date Uploaded
Date Modified
2014-05-01T02:00:07.074+00:00
Audit Status
Audits have not yet been run on this file.
Characterization
File format: pdf (Portable Document Format)
Mime type: application/pdf
File size: 5742494
Last modified: 2015:10:12 20:39:50-06:00
Filename: Sekar_Yokananth_Spring+2011.pdf
Original checksum: 2fe9f8dced34c2155ce2859989dc5a72
Well formed: true
Valid: true
File title: Yoks_Thesis_07Jan_2011.doc
File title: Thesis Front Matter
File author: The University of Calgary
Page count: 258
Activity of users you follow
User Activity Date