title: A transient kinetic analysis of PRMT1 catalysis.
abstract: Post-translational modifications (PTMs) are important strategies used by eukaryotic organisms to modulate their phenotypes. One of the well-studied PTMs, arginine methylation, is catalyzed by protein arginine methyltransferases (PRMTs) with SAM as the methyl donor. The functions of PRMTs have been broadly studied in different biological processes and diseased states, but the molecular basis for arginine methylation is not well-defined. In this study, we report the transient-state kinetic analysis of PRMT1 catalysis. The fast association and dissociation rates suggest that PRMT1 catalysis of histone H4 methylation follows a rapid equilibrium sequential kinetic mechanism. The data give direct evidence that the chemistry of methyl transfer is the major rate-limiting step and that binding of the cofactor SAM or SAH affects the association and dissociation of H4 with PRMT1. Importantly, from the stopped-flow fluorescence measurements, we have identified a critical kinetic step suggesting a precatalytic conformational transition induced by substrate binding. These results provide new insights into the mechanism of arginine methylation and the rational design of PRMT inhibitors.
authors: Department of Chemistry, Georgia State University, Atlanta, Georgia 30302, United States.
authors: You
authors: Y
authors: Feng
authors: Nan
authors: N
authors: Xie
authors: Miyeong
authors: M
authors: Jin
authors: Mary R
authors: MR
authors: Stahley
authors: James T
authors: JT
authors: Stivers
authors: Yujun George
authors: YG
authors: Zheng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D055162:Biocatalysis; D006657:Histones / Q000378:metabolism; D007700:Kinetics; D008745:Methylation; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011484:Protein-Arginine N-Methyltransferases / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D006657:Histones; D010455:Peptides; D011994:Recombinant Proteins; C541817:PRMT1 protein, rat; D011484:Protein-Arginine N-Methyltransferases