Characterization of the universally conserved ATPase YchF using an in vitro and in silico approach

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dc.contributor.authorRosler, Kirsten S
dc.contributor.authorUniversity of Lethbridge. Faculty of Arts and Science
dc.contributor.supervisorWieden, Hans-Joachim
dc.date.accessioned2016-03-08T18:24:17Z
dc.date.available2016-03-08T18:24:17Z
dc.date.issued2014
dc.degree.levelMastersen_US
dc.description.abstractGTPases perform a myriad of functions within the cell, from protein synthesis to cellular signaling. Of all known GTPases, only eight are conserved across all three domains of life. YchF is one of the eight universally conserved GTPases, however its cellular function is poorly understood. YchF differs from the classical GTPases in that it has a higher affinity for ATP than for GTP and functions as an ATPase. As a HAS-GTPase, YchF does not possess the canonical catalytic glutamine required for nucleotide hydrolysis, urging the question of how does YchF hydrolyze ATP. Here we have used molecular dynamics simulations (in silico) and biochemical experiments (in vitro) to identify an amino acid, histidine 114, essential for ATP hydrolysis in YchF. His 114 is located in a flexible loop of the G-domain of YchF, which shows nucleotidedependent conformations in silico. The findings reported indicate that the 70S ribosome can stimulate the ATPase activity of YchF by directly participating in catalysis as well as helping to position the catalytic histidine residue. Additionally, nucleotide binding and dissociation rate constants have been determined in the presence and absence of Mg2+ in order to further understand the functional cycle of YchF.en_US
dc.embargoNoen_US
dc.identifier.urihttps://hdl.handle.net/10133/4442
dc.language.isoen_CAen_US
dc.proquest.subject0487en_US
dc.proquestyesYesen_US
dc.publisherLethbridge, Alta : University of Lethbridge, Dept. of Chemistry and Biochemistryen_US
dc.publisher.departmentDepartment of Chemistry and Biochemistryen_US
dc.publisher.facultyFaculty of Arts and Scienceen_US
dc.relation.ispartofseriesThesis (University of Lethbridge. Faculty of Arts and Science)en_US
dc.subjectYchFen_US
dc.subjectHAS-GTPaseen_US
dc.subjectATPaseen_US
dc.subjectcatalytic mechanismen_US
dc.subjectnucleotide bindingen_US
dc.titleCharacterization of the universally conserved ATPase YchF using an in vitro and in silico approachen_US
dc.typeThesisen_US
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