Type I protein arginine methyltransferases catalyze the formation of asymmetric ω-N G,N G-dimethylarginine residues by transferring methyl groups fromS-adenosyl-l-methionine to guanidino in a variety eucaryotic proteins. The predominant type enzyme activity is found mammalian cells as high molecular weight complex (300–400 kDa). In previous study, this methyltransferase was identified an additional 10-formyltetrahydrofolate dehydrogenase (FDH) protein. However, immunodepletion FDH RAT1 and...

We have identified a new mammalian protein arginine <i>N</i>-methyltransferase, PRMT5, formerly designated Janus kinase-binding 1, that can catalyze the formation of ω-<i>N</i> ^G-monomethylarginine and symmetric ^G,<i>N</i> ^G′-dimethylarginine in variety proteins. A hemagglutinin peptide-tagged PRMT5 complex purified from human HeLa cells catalyzes the<i>S</i>-adenosyl-l-[<i>methyl-</i> ³H]methionine-dependent<i>in vitro</i> methylation myelin basic...

Protein arginine methylation is a prevalent posttranslational modification in eukaryotic cells that has been implicated signal transduction, the metabolism of nascent pre-RNA, and transcriptional activation processes. In searching human genome for protein N-methyltransferase (PRMT) family members, novel gene found on chromosome 1 encodes an apparent methyltransferase, PRMT6. The polypeptide chain PRMT6 41.9 kDa consisting catalytic core sequence common to other PRMT enzymes. Expressed as...

Src homology 3 (SH3) and WW domains are known to associate with proline-rich motifs within their respective ligands. Here we demonstrate that the proposed adapter protein for kinases, Sam68, is a ligand whose interact SH3 of p59 fyn phospholipase Cγ-1 as well FBP30 FBP21. These motifs, in turn, flanked by RG repeats represent targets type I arginine N-methyltransferase. The asymmetrical dimethylation residues these dramatically reduces binding Cγ-1, but has no effect on domain FBP30. results...

We have identified a mammalian arginine N-methyltransferase, PRMT7, that can catalyze the formation of ω-NG-monomethylarginine in peptides. This protein is encoded by gene on human chromosome 16q22.1 (human locus AK001502). expressed full-length cDNA construct Escherichia coli as glutathione S-transferase (GST) fusion protein. found GST-tagged PRMT7 catalyzes S-adenosyl-[methyl-3H]-l-methionine-dependent methylation synthetic peptide GGPGGRGGPGG-NH2 (R1). The radiolabeled was purified...

Protein arginine N-methyltransferases (PRMTs) methylate residues within proteins using S-adenosyl-L-methionine (AdoMet) to form S-adenosyl-L-homocysteine and methylarginine residues. All PRMTs produce ω-NG-monomethylarginine (MMA) either asymmetric ω-NG,NG-dimethylarginine (aDMA) or symmetric ω-NG,N′G-dimethylarginine (sDMA) residues, referred as Type I II activity respectively. Here we report methylation from PRMT2 compare it with PRMT1 UPLC-MS/MS (ultra-performance liquid...

S-Adenosyl-l-methionine-dependent protein arginine N-methyltransferases (PRMTs) catalyze the methylation of residues within a variety proteins. At least four distinct mammalian family members have now been described, including PRMT1, PRMT3, CARM1/PRMT4, and JBP1/PRMT5. To more fully define physiological role we characterized its unique putative zinc-finger domain how it can affect enzymatic activity. Here show that PRMT3 does contain single in amino terminus. Although zinc-liganded form this...

Ubiquinone (coenzyme Q or Q) is a lipid that functions in the electron transport chain inner mitochondrial membrane of eukaryotes and plasma prokaryotes. Q-deficient mutants Saccharomyces cerevisiae harbor defects one eight COQ genes (coq1–coq8) are unable to grow on nonfermentable carbon sources. The biosynthesis involves two separate O-methylation steps. In yeast, first utilizes 3,4-dihydroxy-5-hexaprenylbenzoic acid as substrate thought be catalyzed by Coq3p, 32.7-kDa protein 40%...

Human protein arginine N-methyltransferase 6 (PRMT6) transfers methyl groups from the co-substrate S-adenosyl-L-methionine to residues within proteins, forming S-adenosyl-L-homocysteine as well omega-N(G)-monomethylarginine (MMA) and asymmetric dimethylarginine (aDMA) in process. We have characterized kinetic mechanism of recombinant His-tagged PRMT6 using a mass spectrometry method for monitoring methylation series peptides bearing single arginine, MMA, or aDMA residue. find that follows an...

Protein arginine N-methyltransferases (PRMTs) act in signaling pathways and gene expression by methylating residues within target proteins. PRMT1 is responsible for most cellular methylation activity can work independently or collaboration with other PRMTs. In this study, we demonstrate a direct interaction between PRMT2 using co-immunoprecipitation, bimolecular fluorescence complementation, enzymatic assays. As result of interaction, stimulated activity, affecting its apparent V(max) K(M)...

PRMT6 is a type I protein arginine methyltransferase, generating the asymmetric dimethylarginine mark on proteins such as histone H3R2. Asymmetric dimethylation of H3R2 by acts repressive that antagonizes trimethylation H3 lysine 4 MLL H3K4 methyltransferase. overexpressed in several cancer types, including prostate, bladder and lung cancers; therefore, it great interest to develop potent selective inhibitors for PRMT6. Here, we report synthesis bisubstrate inhibitor GMS [6'-methyleneamine...

Protein arginine N-methyltransferases (PRMTs) catalyze the post-translational methylation of residues within substrate proteins. Their roles in epigenetic regulation gene expression make them viable targets for drug discovery. Peptides containing a single residue substituted at guanidino nitrogen (N(η)) with an ethyl group bearing zero to three fluorine atoms (R1-1, -2, -3, and -4) have been synthesized tested inhibition activity PRMT1, PRMT6, CARM1. Only nonfluorinated R1-1 peptide is...

Double player: Protein arginine N-methyltransferases (PRMTs) employ a general SN2-like bisubstrate reaction mechanism with the cofactor S-adenosyl-L-methionine (AdoMet) to methylate L-arginine residues in target proteins. In this study, new peptidic partial analogues, bearing minimal AdoMet fragment (highlighted yellow) were prepared and evaluated as PRMT inhibitors. Detailed facts of importance specialist readers are published "Supporting Information". Such documents peer-reviewed, but not...

Protein arginine N-methyltransferase 2 (PRMT2) functions in JAK-STAT and Wnt/β-catenin signalling pathways, serves as a nuclear receptor-dependent transcriptional co-activator, represses NF-κB E2F1 transcription factor activities to promote apoptosis. We have previously demonstrated that PRMT2 interacts with PRMT1 increases its activity. Here, we reveal associations using proteomics between the SH3 domain splicing factors including Src-associated mitosis 68 kDa protein (SAM68), substrate...

The HSL7 (histone synthetic lethal 7) gene in the yeast Saccharomyces cerevisiae encodes a protein with close sequence similarity to mammalian PRMT5 protein, member of class arginine methyltransferases that catalyses formation ω-NG-monomethylarginine and symmetric ω-NG,N′G-dimethylarginine residues number methyl-accepting species. A full-length construct was expressed as FLAG-tagged cerevisiae. We found Hsl7 effectively transfer methyl groups from S-adenosyl-[methyl-3H]-L-methionine calf...

Protein arginine N-methyltransferase (PRMT) dimerization is required for methyl group transfer from the cofactor S-adenosyl-L-methionine (AdoMet) to residues in protein substrates, forming S-adenosyl-L-homocysteine (AdoHcy) and methylarginine residues. In this study, we use Förster resonance energy (FRET) determine dissociation constant (K(D)) values of PRMT1 PRMT6. By attaching monomeric Cerulean Citrine fluorescent proteins their N-termini, PRMTs are formed that exhibit similar enzyme...

Abstract Arginine methylation is a prevalent post‐translational modification in eukaryotic cells. Two significant debates exist within the field: do these enzymes dimethylate their substrates processive or distributive manner, and operate using random sequential method of bisubstrate binding? We revealed that human protein arginine N ‐methyltransferase 1 (PRMT1) enzyme kinetics are dependent on substrate sequence. Further, peptides containing an η‐hydroxyarginine generally demonstrated...