Date of Award

2008

Degree Type

Thesis

Degree Name

Master of Science

Program

Microbiology and Immunology

Supervisor

Dr. Carole Creuzenet

Second Advisor

Dr. Wayne Flintoff

Third Advisor

Dr. John McCormick

Abstract

Putative type III secretion system (TTSS)-Iike proteins were identified in silica in H. pylori. These include two cargo proteins, HP0235 and HP1563 (TsaA) and one chaperone, HP1382. The cargo proteins may be secreted via the H. pylori flagellum in a TTSS-like manner. Two experimental approaches were implemented to identify interacting partners of the TTSS-like proteins. First, HP0235 and HP13 82 were TAP- tagged and introduced into H. pylori. HP0235 was co-purified with its associating partners by IgG affinity chromatography, but low protein expression and recovery prevented downstream analyses. Expression of HP1382 was not inducible by any condition known to induce the TTSS. In an alternative approach, the TTSS-like cargo proteins, HP0235 and TsaA, were histidine-tagged and over-expressed in E. coli, loaded onto Ni2+ columns, and used to pull out interacting partners from H. pylori lysates. The detected partners await identification. Also, secretome analyses showed active secretion of HP0235 and TsaA in non-glycosylated forms. Determining the interactions of the TTSS-like proteins should facilitate understanding of their biological role in H. pylori pathogenesis.

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