Expression, purification, crystallization and preliminary crystallographic study of isolated modules of the mouse coactivator-associated arginine methyltransferase 1

Troffer-Charlier, N.; Cura, Vincent; Hassenboehler, P.; Moras, Dino; Cavarelli, Jean

doi:10.1107/s1744309107011785

Cited by 11 publications

(14 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A construct of mouse CARM1 from residues 130 to 487 (mCARM1 130‐487 ) was expressed as a GST fusion protein using the baculovirus system, purified as previously described and crystallized in 15% (w/v) PEG 2000MME, 100 m m Tris/HCl pH 8.0, 100 m m NaCl in the presence of 1 m m Cp1 at 20 °C. mCARM1 crystals were cryoprotected by a short soak in 22% (w/v) PEG 2000MME, 100 m m Tris/HCl pH 8.0, 100 m m NaCl, 15% (v/v) PEG 400, and flash‐cooled in liquid nitrogen.…”

Section: Methodsmentioning

confidence: 99%

Structural studies of protein arginine methyltransferase 2 reveal its interactions with potential substrates and inhibitors

et al. 2016

Self Cite

View full text Add to dashboard Cite

PRMT2 is the less-characterized member of the protein arginine methyltransferase family in terms of structure, activity, and cellular functions. PRMT2 is a modular protein containing a catalytic Ado-Met-binding domain and unique Src homology 3 domain that binds proteins with proline-rich motifs. PRMT2 is involved in a variety of cellular processes and has diverse roles in transcriptional regulation through different mechanisms depending on its binding partners. PRMT2 has been demonstrated to have weak methyltransferase activity on a histone H4 substrate, but its optimal substrates have not yet been identified. To obtain insights into the function and activity of PRMT2, we solve several crystal structures of PRMT2 from two homologs (zebrafish and mouse) in complex with either the methylation product S-adenosyl-L-homocysteine or other compounds including the first synthetic PRMT2 inhibitor (Cp1) studied so far. We reveal that the N-terminal-containing SH3 module is disordered in the full-length crystal structures, and highlights idiosyncratic features of the PRMT2 active site. We identify a new nonhistone protein substrate belonging to the serine-/ arginine-rich protein family which interacts with PRMT2 and we characterize six methylation sites by mass spectrometry. To better understand structural basis for Cp1 binding, we also solve the structure of the complex PRMT4:Cp1. We compare the inhibitor-protein interactions occurring in the PRMT2 and PRMT4 complex crystal structures and show that this compound inhibits efficiently PRMT2. These results are a first step toward a better understanding of PRMT2 substrate recognition and may accelerate the development of structure-based drug design of PRMT2 inhibitors.Database All coordinates and structure factors have been deposited in the Protein Data Bank: zPRMT2 1-408 -SFG = 5g02; zPRMT2 73-408 -SAH = 5fub; mPRMT2 1-445 -SAH = 5ful; mPRMT2 1-445 -Cp1 = 5fwa, mCARM1 130-487 -Cp1 = 5k8v.Abbreviations mPRMT2, mouse (Mus musculus) PRMT2; PDB, Protein Data Bank; PRMT, protein arginine methyltransferase; RMSD, root-square-mean

show abstract

Section: Methodsmentioning

confidence: 99%

Structural studies of protein arginine methyltransferase 2 reveal its interactions with potential substrates and inhibitors

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

“…Details of the expression, purification and crystallization of CARM1 28–140 are given in the Supplementary data. CARM1 28–507 , apo and SAH‐CARM1 140–480 have been overexpressed, purified and crystallized as already published (Troffer‐Charlier et al , 2007).…”

Section: Methodsmentioning

confidence: 99%

Functional insights from structures of coactivator-associated arginine methyltransferase 1 domains

Troffer-Charlier¹,

Cura²,

Hassenboehler³

et al. 2007

EMBO J

Self Cite

137

202

View full text Add to dashboard Cite

Coactivator-associated arginine methyltransferase 1 (CARM1), a protein arginine methyltransferase recruited by several transcription factors, methylates a large variety of proteins and plays a critical role in gene expression. We report, in this paper, four crystal structures of isolated modules of CARM1. The 1.7 Å crystal structure of the N-terminal domain of CARM1 reveals an unexpected PH domain, a scaffold frequently found to regulate protein-protein interactions in a large variety of biological processes. Three crystal structures of the CARM1 catalytic module, two free and one cofactor-bound forms (refined at 2.55 Å , 2.4 Å and 2.2 Å , respectively) reveal large structural modifications including disorder to order transition, helix to strand transition and active site modifications. The N-terminal and the C-terminal end of CARM1 catalytic module contain molecular switches that may inspire how CARM1 regulates its biological activities by protein-protein interactions.

show abstract

“…Next the compounds were screened against selected human PRMTs (table 3), identified to play a role in cancer development and maintenance, and co-crystallization studies were performed with compounds 1, 2, 3, 4 and 7 with an isolated catalytic domain of MmPRMT4 (residues 130 to 487) [46,47] or with PRMT6 [30]. Interestingly, compound 2 and its analogue 4 showed an activity against PRMT4, also called coactivatorassociated arginine methyltransferase 1 (CARM1), and some activity against PRMT7 (table 3).…”

Section: (C) Biological Activitymentioning

confidence: 99%

Hijacking DNA methyltransferase transition state analogues to produce chemical scaffolds for PRMT inhibitors

Halby

Marechal

Pechalrieu

et al. 2018

Phil. Trans. R. Soc. B

Self Cite

View full text Add to dashboard Cite

DNA, RNA and histone methylation is implicated in various human diseases such as cancer or viral infections, playing a major role in cell process regulation, especially in modulation of gene expression. Here we developed a convergent synthetic pathway starting from a protected bromomethylcytosine derivative to synthesize transition state analogues of the DNA methyltransferases. This approach led to seven 5-methylcytosine-adenosine compounds that were, surprisingly, inactive against hDNMT1, hDNMT3Acat, TRDMT1 and other RNA human and viral methyltransferases. Interestingly, compound and its derivative showed an inhibitory activity against PRMT4 in the micromolar range. Crystal structures showed that compound binds to the PRMT4 active site, displacing strongly the-adenosyl-l-methionine cofactor, occupying its binding site, and interacting with the arginine substrate site through the cytosine moiety that probes the space filled by a substrate peptide methylation intermediate. Furthermore, the binding of the compounds induces important structural switches. These findings open new routes for the conception of new potent PRMT4 inhibitors based on the 5-methylcytosine-adenosine scaffold.This article is part of a discussion meeting issue 'Frontiers in epigenetic chemical biology'.

show abstract

Expression, purification, crystallization and preliminary crystallographic study of isolated modules of the mouse coactivator-associated arginine methyltransferase 1

Cited by 11 publications

References 14 publications

Structural studies of protein arginine methyltransferase 2 reveal its interactions with potential substrates and inhibitors

Structural studies of protein arginine methyltransferase 2 reveal its interactions with potential substrates and inhibitors

Functional insights from structures of coactivator-associated arginine methyltransferase 1 domains

Hijacking DNA methyltransferase transition state analogues to produce chemical scaffolds for PRMT inhibitors

Contact Info

Product

Resources

About