Insight into the universally conserved NTPases HflX and YchF

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Coatham, Mackenzie Leigh
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Lethbridge, Alta. : University of Lethbridge, Dept. of Chemistry and Biochemistry
The functional roles of the two universally conserved bacterial GTPases, HflX and YchF, are poorly understood. Both proteins associate with 70S ribosomes as well as 30S and 50S ribosomal subunits. Understanding exactly how HflX and YchF interact with the ribosome and nucleotides will be important for the discovery of the in vivo relevant ribosomal complex. Presented in this thesis, is the development of a fluorescence-based system that can be used to monitor the association of HflX to 70S, 50S and 30S. Additionally, as HflX lacks the canonical glutamine that is required for the hydrolysis of GTP and ATP, an examination into how HflX hydrolyzes purine nucleotides was conducted. Furthermore, nucleotide association and dissociation rate constants were determined in the presence of ribosomes for YchF and in the presence and absence of antibiotics for HflX. The results presented here provide additional insight into the enzymatic properties of HflX and YchF.
bacteria , enzymatic properties , GTPases , HflX , YchF