Crystal structure of arginine methyltransferase 6 from Trypanosoma brucei

Chongyuan Wang; Yuwei Zhu; Jiajia Chen; Xu Li; Junhui Peng; Jiajing Chen; Yang Zou; Zhiyong Zhang; Hong Jin; Pengyuan Yang; Jihui Wu; Liwen Niu; Qingguo Gong; Maikun Teng; Yunyu Shi

doi:10.1371/journal.pone.0087267

Crystal structure of arginine methyltransferase 6 from Trypanosoma brucei

PLoS One. 2014 Feb 3;9(2):e87267. doi: 10.1371/journal.pone.0087267. eCollection 2014.

Authors

Chongyuan Wang¹, Yuwei Zhu¹, Jiajia Chen², Xu Li¹, Junhui Peng¹, Jiajing Chen¹, Yang Zou¹, Zhiyong Zhang¹, Hong Jin², Pengyuan Yang², Jihui Wu¹, Liwen Niu¹, Qingguo Gong¹, Maikun Teng¹, Yunyu Shi¹

Affiliations

¹ Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Anhui, China.
² Department of Chemistry and Institute of Biomedical Science, Fudan University, Shanghai, China.

Abstract

Arginine methylation plays vital roles in the cellular functions of the protozoan Trypanosoma brucei. The T. brucei arginine methyltransferase 6 (TbPRMT6) is a type I arginine methyltransferase homologous to human PRMT6. In this study, we report the crystal structures of apo-TbPRMT6 and its complex with the reaction product S-adenosyl-homocysteine (SAH). The structure of apo-TbPRMT6 displays several features that are different from those of type I PRMTs that were structurally characterized previously, including four stretches of insertion, the absence of strand β15, and a distinct dimerization arm. The comparison of the apo-TbPRMT6 and SAH-TbPRMT6 structures revealed the fine rearrangements in the active site upon SAH binding. The isothermal titration calorimetry results demonstrated that SAH binding greatly increases the affinity of TbPRMT6 to a substrate peptide derived from bovine histone H4. The western blotting and mass spectrometry results revealed that TbPRMT6 methylates bovine histone H4 tail at arginine 3 but cannot methylate several T. brucei histone tails. In summary, our results highlight the structural differences between TbPRMT6 and other type I PRMTs and reveal that the active site rearrangement upon SAH binding is important for the substrate binding of TbPRMT6.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Apoenzymes / chemistry
Apoenzymes / metabolism
Arginine / metabolism
Blotting, Western
Catalytic Domain
Cattle
Crystallography, X-Ray
Histones / metabolism
Mass Spectrometry
Methylation
Models, Molecular
Molecular Sequence Data
Protein Binding
Protein Multimerization
Protein Structure, Tertiary*
Protein-Arginine N-Methyltransferases / chemistry*
Protein-Arginine N-Methyltransferases / genetics
Protein-Arginine N-Methyltransferases / metabolism
Protozoan Proteins / chemistry*
Protozoan Proteins / genetics
Protozoan Proteins / metabolism
S-Adenosylhomocysteine / chemistry
S-Adenosylhomocysteine / metabolism
Sequence Homology, Amino Acid
Substrate Specificity
Trypanosoma brucei brucei / enzymology*
Trypanosoma brucei brucei / genetics
Trypanosoma brucei brucei / metabolism

Substances

Apoenzymes
Histones
Protozoan Proteins
Arginine
S-Adenosylhomocysteine
Protein-Arginine N-Methyltransferases

Grants and funding

Financial support for this project was provided by the Chinese Ministry of Science and Technology (Grant Nos. 2012CB917200, 2012CB917201, 2011CB911104, 2011CB966302 and 2009CB825500), Chinese National Science Foundation (Grant Nos. 31170693, 31330018, 31270782, 30830031, 31270014, 31130018, 31270760 and 30900224), Grant of Chinese Academy of Science (KJZD-EW-L05), Anhui Nature Science Foundation (Grant No. 1208085MC38. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.