Show simple item record

dc.contributor.advisorLoh, Stewart
dc.contributor.authorKarchin, Joshua Michael
dc.date.accessioned2021-07-30T18:34:09Z
dc.date.available2021-07-30T18:34:09Z
dc.date.issued2016
dc.identifier.urihttp://hdl.handle.net/20.500.12648/2018
dc.description.abstractDomain swapping is a mechanism for proteins to form dimers and higher order oligomers through the exchange of a section of their 3D structures. The backbone peptides of domain swapped oligomers are intertwined, but their 3D structures remain identical to their monomeric state, except for where they cross-over, termed a hinged region. We have developed a technique to engineer domain swapping interfaces with mutually exclusive folding (MEF). MEF achieves this by inserting a ‘lever’ protein into the surface loops of a host ‘target’ protein to form a target-lever fusion. This target-leveris conformationally strained with the lever and the target in a thermodynamic tug-of-war. When the lever is folded, the long distance between its N- and C-termini stretches apart the target and splits it in half. Conversely, when the target is folded, the short length of the loop where the lever was inserted compresses the lever and unfolds it. Domain swapping provides an escape from this tug-of-war as it allows the split target to refold and bypass the conformational strain. Because the lever is external to the target, adjusting the stability of the lever, through well-established thermodynamic principles, allows the propensity for domain swapping to be modulated without affecting the binding interface. This enables the design of “triggerable” levers which can reversibly induce domain swapping in response to a signal. Further, we can use domain swapping to turn the function of a target domain on and off. Two target-lever constructs are created with functional mutants in the target domain, one N-terminal to the lever and the other one C-terminal to the lever. Individually, both of these mutants are inactive, however if they are mixed and allowed to domain swap, then up to half of the target domains can swap out the functional mutations into the native active form. This bimolecular system in combination with induced domain swapping enables the design of modular bioswitches and biosensors.en_US
dc.language.isoen_USen_US
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectDesignen_US
dc.subjectCharacterizationen_US
dc.subjectProteinsen_US
dc.subjectRationallyen_US
dc.subjectEngineereden_US
dc.subjectDomainen_US
dc.subjectSwapen_US
dc.subjectMutuallyen_US
dc.subjectExclusiveen_US
dc.subjectFoldingen_US
dc.titleDesign and Characterization of Proteins Rationally Engineered to Domain Swap by Mutually Exclusive Foldingen_US
dc.typeDissertationen_US
dc.description.versionNAen_US
refterms.dateFOA2021-07-30T18:34:10Z
dc.description.institutionUpstate Medical Universityen_US
dc.description.departmentBiochemistry and Molecular Biologyen_US
dc.description.degreelevelPhDen_US
dc.identifier.oclc1006896863


Files in this item

Thumbnail
Name:
Karchin_Josh.pdf
Size:
98.50Mb
Format:
PDF

This item appears in the following Collection(s)

Show simple item record

Attribution-NonCommercial-NoDerivatives 4.0 International
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivatives 4.0 International