Structure and function of PhyA from Desulfovibrio magneticus and use of its product in affinity pull-down experiments

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dc.contributor.authorCleland, Colyn
dc.contributor.authorUniversity of Lethbridge. Faculty of Arts and Science
dc.contributor.supervisorMosimann, Steven
dc.contributor.supervisorSelinger, L. Brent
dc.date.accessioned2019-07-22T17:22:55Z
dc.date.available2019-07-22T17:22:55Z
dc.date.issued2019
dc.degree.levelMastersen_US
dc.description.abstractThe X-ray crystallographic structure of a divergent PTPLP from Desulfovibrio magneticus has been determined in the presence and absence of InsP6 substrate in order to identify the structural features that give rise to its 3-4-5 substrate specificity. These include a novel Phy loop conformation and R242 of the HCRGG.GR P-loop signature sequence. Further, PTPLPs containing an arginine following the cysteine nucleophile are expected to preferentially target the C4-phosphoryl of Ins(1,2,4,5,6)P5 and have a 3-4 hydrolytic pathway. Subsequently, both InsP6 and Ins(1,2,6)P3 were covalently attached to chromatographic resin and utilized as ’bait’ molecules in novel affinity pull-down experiments utilizing a phosphate control column and unfractionated whole cell lysates. The resulting interactomes contain a greater fraction of known IP interacting proteins than current literature methods and identify similar or larger number of total proteins. Despite these improvements, more work needs to be done to address non-specific binding to our columns.en_US
dc.identifier.urihttps://hdl.handle.net/10133/5478
dc.language.isoen_USen_US
dc.proquest.subjectMolecular biology [0307]en_US
dc.proquest.subjectBiochemistry [0487]en_US
dc.proquest.subjectMolecular physics [0609]en_US
dc.proquestyesYesen_US
dc.publisherLethbridge, Alta. : University of Lethbridge, Dept. of Chemistry & Biochemistryen_US
dc.publisher.departmentDepartment of Chemistry & Biochemistryen_US
dc.publisher.facultyArts and Scienceen_US
dc.relation.ispartofseriesThesis (University of Lethbridge. Faculty of Arts and Science)en_US
dc.subjectprotein complexen_US
dc.subjectsubstrate specificityen_US
dc.subjectPTP-likeen_US
dc.subjectaffinity pull-downen_US
dc.subjectCysteine proteinasesen_US
dc.subjectInositol phosphatesen_US
dc.subjectPhosphatasesen_US
dc.subjectX-ray crystallographyen_US
dc.titleStructure and function of PhyA from Desulfovibrio magneticus and use of its product in affinity pull-down experimentsen_US
dc.typeThesisen_US
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