Phe71 in Type III Trypanosomal Protein Arginine Methyltransferase 7 (TbPRMT7) Restricts the Enzyme to Monomethylation

Tamar B. Cáceres, Abhishek Thakur, Owen M. Price, Nicole Ippolito, Jun Li, Jun Qu, Orlando Acevedo, Joan M. Hevel

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Protein arginine methyltransferase 7 (PRMT7) is unique within the PRMT family as it is the only isoform known to exclusively make monomethylarginine (MMA). Given its role in epigenetics, the mechanistic basis for the strict monomethylation activity is under investigation. It is thought that PRMT7 enzymes are unable to add a second methyl group because of steric hindrance in the active site that restricts them to monomethylation. To test this, we probed the active site of trypanosomal PRMT7 (TbPRMT7) using accelerated molecular dynamics, site-directed mutagenesis, kinetic, binding, and product analyses. Both the dynamics simulations and experimental results show that the mutation of Phe71 to Ile converts the enzyme from a type III methyltransferase into a mixed type I/II, that is, an enzyme that can now perform dimethylation. In contrast, the serine and alanine mutants of Phe71 preserve the type III behavior of the native enzyme. These results are inconsistent with a sterics-only model to explain product specificity. Instead, molecular dynamics simulations of these variants bound to peptides show hydrogen bonding between would-be substrates and Glu172 of TbPRMT7. Only in the case of the Phe71 to Ile mutation is this interaction between MMA and the enzyme maintained, and the geometry for optimal SN2 methyl transfer is obtained. The results of these studies highlight the benefit of combined computational and experimental methods in providing a better understanding for how product specificity is dictated by PRMTs.

Original languageEnglish (US)
Pages (from-to)1349-1359
Number of pages11
JournalBiochemistry
Volume57
Issue number8
DOIs
StatePublished - Feb 27 2018

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Protein-Arginine N-Methyltransferases
Enzymes
Molecular Dynamics Simulation
Molecular dynamics
Catalytic Domain
Mutagenesis
Mutation
Computer simulation
Methyltransferases
Hydrogen Bonding
Site-Directed Mutagenesis
Epigenomics
Alanine
Serine
Hydrogen bonds
Protein Isoforms
Peptides
Kinetics
Geometry
Substrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Cáceres, T. B., Thakur, A., Price, O. M., Ippolito, N., Li, J., Qu, J., ... Hevel, J. M. (2018). Phe71 in Type III Trypanosomal Protein Arginine Methyltransferase 7 (TbPRMT7) Restricts the Enzyme to Monomethylation. Biochemistry, 57(8), 1349-1359. https://doi.org/10.1021/acs.biochem.7b01265

Phe71 in Type III Trypanosomal Protein Arginine Methyltransferase 7 (TbPRMT7) Restricts the Enzyme to Monomethylation. / Cáceres, Tamar B.; Thakur, Abhishek; Price, Owen M.; Ippolito, Nicole; Li, Jun; Qu, Jun; Acevedo, Orlando; Hevel, Joan M.

In: Biochemistry, Vol. 57, No. 8, 27.02.2018, p. 1349-1359.

Research output: Contribution to journalArticle

Cáceres, Tamar B. ; Thakur, Abhishek ; Price, Owen M. ; Ippolito, Nicole ; Li, Jun ; Qu, Jun ; Acevedo, Orlando ; Hevel, Joan M. / Phe71 in Type III Trypanosomal Protein Arginine Methyltransferase 7 (TbPRMT7) Restricts the Enzyme to Monomethylation. In: Biochemistry. 2018 ; Vol. 57, No. 8. pp. 1349-1359.
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