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Author ORCID Identifier
https://orcid.org/0000-0001-7952-1551
AccessType
Open Access Dissertation
Document Type
dissertation
Degree Name
Doctor of Philosophy (PhD)
Degree Program
Chemistry
Year Degree Awarded
2020
Month Degree Awarded
September
First Advisor
Eric Strieter
Subject Categories
Chemistry | Life Sciences
Abstract
Ubiquitin is a 76 amino acids protein that is evolutionary conserved in eukaryotes. It is an important signaling molecule in a plethora biological events, such as protein degradation, DNA damage response, and transcription. This thesis aims to develop engineered protein as a tool to study ubiquitin signaling. Through targeted mutagenesis and directed evolution, a deubiquitinase is reprogrammed into a transamidase, which lead to the generation of ubiquitinprotein conjugates with discrete ubiquitin linkages through auto-ubiquitination. These ubiquitin-protein conjugates could be used as a model substrate to profile their interaction of different ubiquitin interacting proteins. In addition, using directed evolution and deep sequencing, a nanobody-based binder targeting a deubiquitinase is engineered. This nanobody could be utilized as an intracellular probe to study the regulation of the deubiquitinase with great spatial and temporal control.
DOI
https://doi.org/10.7275/18414115
Recommended Citation
Chang, Lin Hui, "Engineered proteins as tools to understand ubiquitin signaling" (2020). Doctoral Dissertations. 2001.
https://doi.org/10.7275/18414115
https://scholarworks.umass.edu/dissertations_2/2001