Class
Article
College
College of Science
Department
English Department
Faculty Mentor
Joan Hevel
Presentation Type
Oral Presentation
Abstract
Protein arginine methyltransferase 1 (PRMT1) is an enzyme that post-translationally methylates other proteins in the cell. These methylation events play critical roles in maintaining cellular health. Labeling PRMT1 site-specifically will enable further research in PRMT1 oligomeric state in vivo and can be applied in future fluorescent studies. The intention of this research was to identify a position to label PRMT1 that does not affect oligomerization or activity. Position 312 was chosen as the site for specific tagging as it will have less interference in oligomerization, minimizes free rotation of fluorescent tags, and is surface-exposed. To confirm that position 312 was a justifiable position for labeling, a methyltransferase activity of E312C PRMT1 was conducted before and after fluorescent labeling with Alexa Fluor 488—a thiol reactive dye. Native-PAGE with Western blot analysis was also applied to determine oligomeric state of labeled and unlabeled E312C PRMT1. Findings show unlabeled and labeled E312C PRMT1 had similar rates of activity, and oligomeric state of unlabeled and labeled E312C PRMT1 resemble oligomeric state of wild-type PRMT1.
Location
Logan, UT
Start Date
4-8-2022 12:00 AM
Included in
Investigating Position 312 in PRMT1 for Future Fluorescent Labeling Studies
Logan, UT
Protein arginine methyltransferase 1 (PRMT1) is an enzyme that post-translationally methylates other proteins in the cell. These methylation events play critical roles in maintaining cellular health. Labeling PRMT1 site-specifically will enable further research in PRMT1 oligomeric state in vivo and can be applied in future fluorescent studies. The intention of this research was to identify a position to label PRMT1 that does not affect oligomerization or activity. Position 312 was chosen as the site for specific tagging as it will have less interference in oligomerization, minimizes free rotation of fluorescent tags, and is surface-exposed. To confirm that position 312 was a justifiable position for labeling, a methyltransferase activity of E312C PRMT1 was conducted before and after fluorescent labeling with Alexa Fluor 488—a thiol reactive dye. Native-PAGE with Western blot analysis was also applied to determine oligomeric state of labeled and unlabeled E312C PRMT1. Findings show unlabeled and labeled E312C PRMT1 had similar rates of activity, and oligomeric state of unlabeled and labeled E312C PRMT1 resemble oligomeric state of wild-type PRMT1.