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Biochemical Characterization of the Putative Glucosyltransferase, WapB, Which Acts on the Uncapped Core Oligosaccharide of Pseudomonas aeruginosa

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dc.contributor.advisor Kimber, Matthew
dc.contributor.author El-Osta, Lana
dc.date.accessioned 2018-09-05T16:08:31Z
dc.date.available 2018-09-05T16:08:31Z
dc.date.copyright 2018-08-27
dc.date.created 2018-08-27
dc.date.issued 2018-09-05
dc.identifier.uri http://hdl.handle.net/10214/14205
dc.description.abstract Pseudomonas aeruginosa is a Gram-negative, opportunistic pathogen known to cause disease in Cystic Fibrosis patients. Lipopolysaccharide (LPS) forms the outermost layer of the outer membrane and is an important virulence factor. LPS is divided conceptually into three domains: lipid A, core oligosaccharide, and (sometimes) O-antigen. Uncapped (O-antigen free) core is often modified with a terminal glucose (GlcIV) on L-RhaA; while a wapB mutant produces LPS lacking GlcIV. At present, little is known regarding the enzymatic properties of WapB. My hypothesis is that WapB is a β-1,2-glucosyltransferase that transfers GlcIV to the uncapped core OS of P. aeruginosa PAO1. WapB proved to be insoluble when expressed, and several solubilization strategies were explored. Of these strategies, membrane extraction using the detergent DDM proved the most promising, suggesting WapB interacts tightly with the membrane. Complementation was also used to show that minimum 20 C-terminal residues and a DXD motif are required for function. en_US
dc.description.sponsorship CIHR, NSERC, Glyconet en_US
dc.language.iso en en_US
dc.subject biochemistry en_US
dc.subject biophysics en_US
dc.subject bioinformatics en_US
dc.subject microbiology en_US
dc.subject Pseudomonas aeruginosa en_US
dc.subject lipopolysaccharide en_US
dc.subject glycosyltransferase en_US
dc.subject WapB en_US
dc.subject glucosyltransferase en_US
dc.subject core oligosaccharide en_US
dc.subject biosynthesis en_US
dc.subject outer core oligosaccharide en_US
dc.subject uncapped glycoform en_US
dc.subject PAO1 en_US
dc.subject MigA en_US
dc.subject WapR en_US
dc.subject rhamnosyltransferase en_US
dc.title Biochemical Characterization of the Putative Glucosyltransferase, WapB, Which Acts on the Uncapped Core Oligosaccharide of Pseudomonas aeruginosa en_US
dc.type Thesis en_US
dc.degree.programme Molecular and Cellular Biology en_US
dc.degree.name Master of Science en_US
dc.degree.department Department of Molecular and Cellular Biology en_US


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