Defining Topologically Oriented Transmembrane Helix Contacts in Integral Membrane Protein EmrE by a Modified Two-Hybrid Approach

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atmire.migration.oldid1983
dc.contributor.advisorTurner, Raymond Joseph
dc.contributor.authorBurt, Jason, Campbell
dc.date.accessioned2014-04-01T17:23:04Z
dc.date.available2014-06-16T07:00:26Z
dc.date.issued2014-04-01
dc.date.submitted2014en
dc.description.abstractStudy of integral membrane proteins is comparatively difficult in comparison to soluble proteins, requiring more specialized techniques. Here is presented an application of the Bacterial Adenylate Cyclase Two-Hybrid (BACTH) method to study oligomerization in membrane proteins applied to model integral membrane multidrug transporter EmrE. The system was unable to observe dimerization between full length monomers of EmrE, likely due to the preferential antiparallel nature of EmrE dimers. Further studies required large numbers of samples, so the Miller assay was adapted to work in a small volume microtitre plate format. Using this new higher throughput assay, constructs of individual EmrE transmembrane helices (TMH) were utilized to perform contact mapping of EmrE dimer structure. The results were compiled to construct a transmembrane helix interaction structure. This structure obtained is novel and identifies inconsistencies from previous studies.en_US
dc.identifier.citationBurt, J. (2014). Defining Topologically Oriented Transmembrane Helix Contacts in Integral Membrane Protein EmrE by a Modified Two-Hybrid Approach (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca. doi:10.11575/PRISM/25539en_US
dc.identifier.doihttp://dx.doi.org/10.11575/PRISM/25539
dc.identifier.urihttp://hdl.handle.net/11023/1397
dc.language.isoeng
dc.publisher.facultyGraduate Studies
dc.publisher.institutionUniversity of Calgaryen
dc.publisher.placeCalgaryen
dc.rightsUniversity of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.
dc.subjectBiochemistry
dc.subject.classificationEmrEen_US
dc.subject.classificationIntegral Membrane Proteinen_US
dc.subject.classificationProtein Structureen_US
dc.subject.classificationTransmembrane Helicesen_US
dc.subject.classificationBacterial Two-Hybriden_US
dc.subject.classificationMiller Assayen_US
dc.titleDefining Topologically Oriented Transmembrane Helix Contacts in Integral Membrane Protein EmrE by a Modified Two-Hybrid Approach
dc.typemaster thesis
thesis.degree.disciplineBiological Sciences
thesis.degree.grantorUniversity of Calgary
thesis.degree.nameMaster of Science (MSc)
ucalgary.item.requestcopytrue
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