Skip to Content
Merck
  • Automethylation of protein arginine methyltransferase 8 (PRMT8) regulates activity by impeding S-adenosylmethionine sensitivity.

Automethylation of protein arginine methyltransferase 8 (PRMT8) regulates activity by impeding S-adenosylmethionine sensitivity.

The Journal of biological chemistry (2013-08-16)
Myles B C Dillon, Heather L Rust, Paul R Thompson, Kerri A Mowen
ABSTRACT

Protein arginine methyltransferase (PRMT) 8 is unique among the PRMTs, as it has a highly restricted tissue expression pattern and an N terminus that contains two automethylation sites and a myristoylation site. PRMTs catalyze the transfer of a methyl group from S-adenosylmethionine (AdoMet) to a peptidylarginine on a protein substrate. Currently, the physiological roles, regulation, and cellular substrates of PRMT8 are poorly understood. However, a thorough understanding of PRMT8 kinetics should provide insights into each of these areas, thereby enhancing our understanding of this unique enzyme. In this study, we determined how automethylation regulates the enzymatic activity of PRMT8. We found that preventing automethylation with lysine mutations (preserving the positive charge of the residue) increased the turnover rate and decreased the Km of AdoMet but did not affect the Km of the protein substrate. In contrast, mimicking automethylation with phenylalanine (i.e. mimicking the increased hydrophobicity) decreased the turnover rate. The inhibitory effect of the PRMT8 N terminus could be transferred to PRMT1 by creating a chimeric protein containing the N terminus of PRMT8 fused to PRMT1. Thus, automethylation of the N terminus likely regulates PRMT8 activity by decreasing the affinity of the enzyme for AdoMet.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-PRMT1 Antibody, Upstate®, from rabbit
Sigma-Aldrich
OverExpress C41(DE3) Chemically Competent Cells, for the highest protein expression
Sigma-Aldrich
OverExpress C41(DE3) pLysS Chemically Competent Cells, for the highest protein expression
Supelco
L-Phenylalanine, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Supelco
L-Phenylalanine, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
L-Phenylalanine, BioUltra, ≥99.0% (NT)
Sigma-Aldrich
L-Phenylalanine, 99%, FCC
Sigma-Aldrich
L-Phenylalanine, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
L-Phenylalanine, reagent grade, ≥98%