Impact of Point Mutations On The Structure and Function of VRK1
Presenter(s)
Leo Doan, Elijah Murray
Abstract
Vaccinia-related kinase 1 (VRK1) is a serine/threonine kinase protein involved in cell cycle regulation, DNA damage response, and chromatin remodeling. VRK1’s structure contains an N-terminal domain for nuclear localization, a catalytic domain for ATP binding and substrate phosphorylation, and regulatory regions for protein interactions. Mutations of the VRK1 catalytic domain, including L195V and A66G, were studied and compared to the wild-type (WT) characterizations. These mutations have been previously linked to genetic disease. A plasmid encoding the His-tagged protein catalytic domain was introduced to BL21 DE3 RIL cells, then cultured. The protein was separated using affinity, desalting, and gel filtration chromatography. Procedures were adapted to the department’s new NSF-funded fast protein liquid chromatography (FPLC) instrument. Circular dichroism (CD) spectroscopy and differential scanning fluorimetry (DSF) were used to study protein structure and stability of the WT and mutant proteins. The binding of nucleotide and two inhibitors (PIK-75 HCl and BI-D1870) was also investigated using DSF. Binding to ligands stabilizes the protein and increases the melting point. Understanding how these mutations affect VRK1’s ligand binding may improve our understanding of this protein and its role in disease. This abstract and poster were also presented at the 2026 ASBMB National Meeting.
College
College of Science & Engineering
Department
Chemistry
Campus
Winona
First Advisor/Mentor
Emily Ruff
Location
Kryzsko Great River Ballroom, Winona, Minnesota; United States
Start Date
4-23-2026 2:00 PM
End Date
4-23-2026 3:00 PM
Presentation Type
Poster Session
Format of Presentation or Performance
In-Person
Session
2b=2pm-3pm
Poster Number
16
Impact of Point Mutations On The Structure and Function of VRK1
Kryzsko Great River Ballroom, Winona, Minnesota; United States
Vaccinia-related kinase 1 (VRK1) is a serine/threonine kinase protein involved in cell cycle regulation, DNA damage response, and chromatin remodeling. VRK1’s structure contains an N-terminal domain for nuclear localization, a catalytic domain for ATP binding and substrate phosphorylation, and regulatory regions for protein interactions. Mutations of the VRK1 catalytic domain, including L195V and A66G, were studied and compared to the wild-type (WT) characterizations. These mutations have been previously linked to genetic disease. A plasmid encoding the His-tagged protein catalytic domain was introduced to BL21 DE3 RIL cells, then cultured. The protein was separated using affinity, desalting, and gel filtration chromatography. Procedures were adapted to the department’s new NSF-funded fast protein liquid chromatography (FPLC) instrument. Circular dichroism (CD) spectroscopy and differential scanning fluorimetry (DSF) were used to study protein structure and stability of the WT and mutant proteins. The binding of nucleotide and two inhibitors (PIK-75 HCl and BI-D1870) was also investigated using DSF. Binding to ligands stabilizes the protein and increases the melting point. Understanding how these mutations affect VRK1’s ligand binding may improve our understanding of this protein and its role in disease. This abstract and poster were also presented at the 2026 ASBMB National Meeting.
Comments
Doan, Leo; Murray, Elijah J