Class
Article
College
College of Science
Department
Chemistry and Biochemistry Department
Faculty Mentor
Joanie Hevel
Presentation Type
Poster Presentation
Abstract
Protein arginine methyltransferase 1 (PRMT1) catalyzes the transfer of a methyl group from S-adenosyl-L-methionine (AdoMet) to specific arginine residues of a target protein. This type of reaction can impact the activity of the target protein and is an important regulatory process in human cells. Dysregulation of PRMT1 has been associated with severe disease in humans, including cancer. Accurate data concerning PRMT1 activity is important to the characterization of the enzyme. The activity of PRMT1 is studied by combining the enzyme with AdoMet and a target protein for methylation and measuring the amount of methylation that occurs over time. AdoMet that can be purchased commercially is understood to be impure, which may lead to bias in data collected using the reagent. AdoMet can also racemize to an inactive stereoisomer, further decreasing the purity of the reagent. To test if the impurities found in commercially available AdoMet affect measured PRMT1 activity, AdoMet was enzymatically synthesized and purified via fast protein liquid chromatography (FPLC) and high pressure liquid chromatography (HPLC). To estimate the usable lifespan of synthesized and purified AdoMet in the lab, a sample of the reagent was subjected to freeze/thaw cycles and analyzed via HPLC. The synthesized reagent was found to be stable through at least 20 freeze/thaw cycles. The activity of PRMT1 using purified AdoMet was compared to the activity of the enzyme using the commercial reagent. The collected data indicate that the impurities found in the commercially available reagent do not significantly affect PRMT1 activity.Presentation Time: Thursday, 10-11 a.m.Zoom link: https://usu-edu.zoom.us/j/87913811390?pwd=RTR2dEJjaEJ1akJVYTdFMmZIcDNrQT09
Location
Logan, UT
Start Date
4-8-2021 12:00 AM
Included in
Substrate Purity and its Effect on PRMT1 Activity
Logan, UT
Protein arginine methyltransferase 1 (PRMT1) catalyzes the transfer of a methyl group from S-adenosyl-L-methionine (AdoMet) to specific arginine residues of a target protein. This type of reaction can impact the activity of the target protein and is an important regulatory process in human cells. Dysregulation of PRMT1 has been associated with severe disease in humans, including cancer. Accurate data concerning PRMT1 activity is important to the characterization of the enzyme. The activity of PRMT1 is studied by combining the enzyme with AdoMet and a target protein for methylation and measuring the amount of methylation that occurs over time. AdoMet that can be purchased commercially is understood to be impure, which may lead to bias in data collected using the reagent. AdoMet can also racemize to an inactive stereoisomer, further decreasing the purity of the reagent. To test if the impurities found in commercially available AdoMet affect measured PRMT1 activity, AdoMet was enzymatically synthesized and purified via fast protein liquid chromatography (FPLC) and high pressure liquid chromatography (HPLC). To estimate the usable lifespan of synthesized and purified AdoMet in the lab, a sample of the reagent was subjected to freeze/thaw cycles and analyzed via HPLC. The synthesized reagent was found to be stable through at least 20 freeze/thaw cycles. The activity of PRMT1 using purified AdoMet was compared to the activity of the enzyme using the commercial reagent. The collected data indicate that the impurities found in the commercially available reagent do not significantly affect PRMT1 activity.Presentation Time: Thursday, 10-11 a.m.Zoom link: https://usu-edu.zoom.us/j/87913811390?pwd=RTR2dEJjaEJ1akJVYTdFMmZIcDNrQT09