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Permanent link (DOI): https://doi.org/10.7939/R3H677

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Carbohydrate-Protein Interactions Studied Using Electrospray Ionization Mass Spectrometry Open Access

Descriptions

Other title
Subject/Keyword
ESI-MS
Quantitative assay
Noncovalent interactions
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Lin, Hong
Supervisor and department
Klassen, John (Chemistry)
Examining committee member and department
McDermott, Mark (Chemistry)
Petersen, Nils (Chemistry)
Department
Department of Chemistry
Specialization

Date accepted
2013-08-23T14:57:20Z
Graduation date
2013-11
Degree
Master of Science
Degree level
Master's
Abstract
This thesis describes the studying of electrospray ionization (ESI) process and the application of the direct ESI-MS assay to study noncovalent carbohydrate-protein interactions in vitro. Protein unfolding induced by ESI process was first investigated. It is proposed that the Coulombic repulsion between the negatively charged residues and liquid/droplet surface charge under certain ESI conditions can induce unfolding of acidic proteins. A deleterious non-uniform response factors phenomenon induced by high molecular weight molecules and complexes in the application of direct ESI-MS assay was also investigated. It is possibly due to the reduction in the number of available surface sites in the ESI droplets upon introduction of large solute and increased competition between protein and the more hydrophilic carbohydrate-protein complex for these sites. Direct ESI-MS assay was also utilized to investigate the stepwise binding of the GM1 pentasaccharide (GM1os) to the cholera toxin B subunit homopentamer and to elucidate positive binding cooperativity.
Language
English
DOI
doi:10.7939/R3H677
Rights
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 these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before 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
Lin, H.; Kitova, E. N.; Johnson, M. A.; Eugenio, L; Ng, K. K. S.; Klassen, J. S. J. Am. Soc. Mass Spectrom. 2012, 23, 2122-2131.Lin, H.; Kitova, E. N.; Klassen, J. S. J. Am. Soc. Mass Spectrom. 2013, Accepted.

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