Rapid kinetic studies of PhyA from Selenomonas ruminantium, and a simplified means of production of a phosphate biosensor

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dc.contributor.authorSmith, Dustin D.
dc.contributor.authorUniversity of Lethbridge. Faculty of Arts and Science
dc.contributor.supervisorSelinger, L. Brent
dc.contributor.supervisorWieden, Hans-Joachim
dc.date.accessioned2017-01-17T20:45:18Z
dc.date.available2017-01-17T20:45:18Z
dc.date.issued2016
dc.degree.levelMastersen_US
dc.description.abstractMyo-inositol polyphosphates (IPs) are ubiquitous in nature and involved in various cellular events. Dephosphorylation of IPs by protein tyrosine phosphatase-like polyphosphatases (PTPLPs) occurs via a complex pathway, and the in vivo function of many of these enzymes remains unknown. In order to further our understanding of PTPLP catalyzed dephosphorylation of IPs; I present rapid kinetics studies of the representative PTPLP PhyA from Selenomonas ruminantium (PhyAsr). These studies revealed kinetic parameters of PhyAsr dimerization, and myo-inositol hexakisphosphate (IP6) binding to the homodimer. In addition to studying PhyAsr, I have developed a simplified methodology to produce a biosensor capable of detecting phosphate release in real-time. The phosphate biosensor is fluorescently labeled and utilizes Escherichia coli Phosphate-binding protein (PhoS). The results show that my proposed methodology yields a functional biosensor and is feasible for large-scale production. I envision this methodology to be versatile and useful for a large number of research applications where detection of free phosphate is required.en_US
dc.description.sponsorshipUniversity of Lethbridgeen_US
dc.embargoNoen_US
dc.identifier.urihttps://hdl.handle.net/10133/4763
dc.language.isoen_USen_US
dc.proquest.subject0487en_US
dc.proquest.subject0786en_US
dc.proquest.subject0306en_US
dc.proquestyesYesen_US
dc.publisherLethbridge, Alta : University of Lethbridge, Dept. of Chemistry and Biochemistryen_US
dc.publisher.departmentDepartment of Biological Sciencesen_US
dc.publisher.departmentDepartment of Chemistry and Biochemistryen_US
dc.publisher.facultyArts and Scienceen_US
dc.relation.ispartofseriesThesis (University of Lethbridge. Faculty of Arts and Science)en_US
dc.subjectBiochemistryen_US
dc.subjectEnzyme kineticsen_US
dc.subjectPhosphate sensoren_US
dc.subjectPTPLPen_US
dc.subjectmyo-inositolen_US
dc.subjectPhosphataseen_US
dc.subjectStopped-flowen_US
dc.subjectPhyAen_US
dc.subjectPhoSen_US
dc.subjectPhosphate releaseen_US
dc.subjectPhosphate binding proteinen_US
dc.subjectPhytaseen_US
dc.titleRapid kinetic studies of PhyA from Selenomonas ruminantium, and a simplified means of production of a phosphate biosensoren_US
dc.typeThesisen_US
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