The role of the diphthamide-containing loop in ADP-ribosylation of eukaryotic elongation factor 2

dc.contributor.advisorMerrill, A. Rod
dc.contributor.authorZhang, Yong
dc.date.accessioned2021-03-31T15:37:30Z
dc.date.available2021-03-31T15:37:30Z
dc.date.copyright2006
dc.degree.departmentDepartment of Molecular Biology and Geneticsen_US
dc.degree.grantorUniversity of Guelphen_US
dc.degree.nameMaster of Scienceen_US
dc.description.abstractEukaryotic elongation factor 2 (eEF2) contains a post-translationally modified histidine residue, referred to as diphthamide, which is the specific ribosylation target of exotoxin A (ETA) secreted by 'Pseudomonas aeruginosa. ' In yeast eEF2, the diphthamide-containing loop (Leu-693-Gly-703) is located at the tip of domain IV. To elucidate the role of the diphthamide-containing loop in the ADP-ribosylation of eEF2, the conserved residues within this loop were replaced with Ala through the use of site-directed mutagenesis. The structural integrity and ADP-ribose acceptor activities of the purified mutant eEF2 proteins were characterized by a variety of fluorescence-based approaches. The single-Ala mutations in the diphthamide-containing loop produced eEF2 proteins with extremely low ADP-ribose acceptor activities, which indicated that this loop plays an important role in the ADP-ribosylation catalyzed by ETA. In addition, the naked imidazole ring of Diph-699 is essential for the acceptance of the ADP-ribose moiety during the toxin-catalyzed reaction.en_US
dc.identifier.urihttps://hdl.handle.net/10214/24285
dc.language.isoen
dc.publisherUniversity of Guelphen_US
dc.rights.licenseAll items in the Atrium are protected by copyright with all rights reserved unless otherwise indicated.
dc.subjecteukaryotic elongation factor 2en_US
dc.subjectdiphthamideen_US
dc.subjectexotoxin Aen_US
dc.subjectPseudomonas aeruginosaen_US
dc.subjectADP-ribosylationen_US
dc.subjectdiphthamide-containing loopen_US
dc.titleThe role of the diphthamide-containing loop in ADP-ribosylation of eukaryotic elongation factor 2en_US
dc.typeThesisen_US

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