Abstract
VRK1 is a vaccinia-related serine/threonine kinase that contributes to the regulation of cell proliferation. Point mutations within the VRK1 sequence are associated with a variety of complex neurodegenerative disorders which previous research suggests are due to changes in kinase activity and/or structure and stability. Point mutations seen with VRK1 may affect how a tail on the C-terminus of the protein interacts with the protein's active site, possibly altering the ability of the protein to bind to specific substrates. This study looked specifically at the point mutation D263G and its effect on protein stability and ability to bind substrates at its active site. The mutation was produced in a His-tagged VRK1 construct plasmid which was then transformed into E. coli bacteria cells, allowing for the purification of the specific mutated VRK1 protein. Experiments were then carried out to determine both the stability of the protein and its substrate binding properties. The stability of the protein was analyzed using circular dichroism and the protein's binding ability was evaluated using differential scanning fluorimetry with ADP and ADP-competitive inhibitors. Results were compared to the wild type VRK1 protein. These experiments are also being extended to other mutations including R321C, which is seen in some patients diagnosed with neuromuscular diseases.
College
College of Science & Engineering
Department
Chemistry
Campus
Winona
First Advisor/Mentor
Emily Ruff
Start Date
4-19-2023 2:00 PM
End Date
4-19-2023 3:00 PM
Presentation Type
Poster Session
Format of Presentation or Performance
In-Person
Session
2b=2pm-3pm
Poster Number
5
Included in
Exploration of Known Disease-Causing VRK1 Mutants
VRK1 is a vaccinia-related serine/threonine kinase that contributes to the regulation of cell proliferation. Point mutations within the VRK1 sequence are associated with a variety of complex neurodegenerative disorders which previous research suggests are due to changes in kinase activity and/or structure and stability. Point mutations seen with VRK1 may affect how a tail on the C-terminus of the protein interacts with the protein's active site, possibly altering the ability of the protein to bind to specific substrates. This study looked specifically at the point mutation D263G and its effect on protein stability and ability to bind substrates at its active site. The mutation was produced in a His-tagged VRK1 construct plasmid which was then transformed into E. coli bacteria cells, allowing for the purification of the specific mutated VRK1 protein. Experiments were then carried out to determine both the stability of the protein and its substrate binding properties. The stability of the protein was analyzed using circular dichroism and the protein's binding ability was evaluated using differential scanning fluorimetry with ADP and ADP-competitive inhibitors. Results were compared to the wild type VRK1 protein. These experiments are also being extended to other mutations including R321C, which is seen in some patients diagnosed with neuromuscular diseases.
