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Author ORCID Identifier
Open Access Dissertation
Doctor of Philosophy (PhD)
Year Degree Awarded
Month Degree Awarded
Chemistry | Life Sciences
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.
Chang, Lin Hui, "Engineered proteins as tools to understand ubiquitin signaling" (2020). Doctoral Dissertations. 2001.