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dc.contributor.authorBrannon, Mary Katherineen_US
dc.date.accessioned2016-12-24T07:00:19Z
dc.date.available2016-12-24T07:00:19Z
dc.date.issued2015-07-02en_US
dc.identifier.othervt_gsexam:5305en_US
dc.identifier.urihttp://hdl.handle.net/10919/73814
dc.description.abstractAdaptor proteins, like Tollip and Tom1, facilitate cellular cargo sorting through their ubiquitin-binding domains. Tollip and Tom1 bind to each other through their TBD and GAT domains, respectively, whereas Tollip interacts with phosphatidylinositol-3-phosphate (PtdIns(3)P)-containing endosomal membranes. Tom1 and Tollip interaction and association with endosomes is proposed to be involved in the lysosomal degradation of polyubiquitinated cargo. Through cellular, biochemical, and biophysical techniques, we have further characterized the association of Tom1 with Tollip. Mutations in the binding interface of the Tom1 GAT and Tollip TBD complex leads to a subcellular mis-localization of both proteins, indicating that Tom1 may serve to direct Tollip to specific cellular pathways. It was determined that Tom1 inhibits the binding of Tollip to PtdIns(3)P and inhibition was reversed when mutations in the binding interface of the Tom1 GAT and Tollip TBD were present. Furthermore, it was established that, upon the binding of Tollip TBD to Tom1 GAT, ubiquitin is inhibited from binding to Tom1 GAT. It was also demonstrated that Tom1 GAT, but not Tollip TBD, can weakly bind to PtdIns(3)P. Consequently, we propose that association of Tom1 may serve to direct Tollip for involvement in specific cell signaling pathways. Gaining insight into the function of Tom1 and Tollip may lead to their use as therapeutic targets for increasing the efficiency of cargo trafficking and also for patients recovering from various cardiac injuries.en_US
dc.format.mediumETDen_US
dc.publisherVirginia Techen_US
dc.rightsThis Item is protected by copyright and/or related rights. Some uses of this Item may be deemed fair and permitted by law even without permission from the rights holder(s), or the rights holder(s) may have licensed the work for use under certain conditions. For other uses you need to obtain permission from the rights holder(s).en_US
dc.subjectTollipen_US
dc.subjectTom1en_US
dc.subjectendosomal traffickingen_US
dc.subjectphosphatidylinositol-3-phosphateen_US
dc.subjectcellular immunofluorescenceen_US
dc.subjectanalytical ultracentrifugationen_US
dc.subjectsurface plasmon resonanceen_US
dc.subjectnuclear magnetic resonanceen_US
dc.titleBinding properties of adaptor proteins Tollip and Tom1en_US
dc.typeThesisen_US
dc.contributor.departmentBiological Sciencesen_US
dc.description.degreeMaster of Scienceen_US
thesis.degree.nameMaster of Scienceen_US
thesis.degree.levelmastersen_US
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen_US
thesis.degree.disciplineBiological Sciencesen_US
dc.contributor.committeechairCapelluto, Daniel G. S.en_US
dc.contributor.committeememberKelly, Deborah F.en_US
dc.contributor.committeememberBevan, David R.en_US
dc.contributor.committeememberSible, Jill C.en_US


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