Myelin Basic Protein Citrullination, a Hallmark of Central Nervous System Demyelination, Assessed by Novel Monoclonal Antibodies in Prion Diseases

  • Byungki Jang
  • Yong-Chul Jeon
  • Hae-Young Shin
  • Yun-Jung Lee
  • Hyunji Kim
  • Yoshitaka Kondo
  • Akihito Ishigami
  • Yong-Sun Kim
  • Eun-Kyoung Choi
Article

Abstract

Myelin basic protein (MBP) citrullination by peptidylarginine deiminase (PAD) enzymes leads to incomplete protein-lipid bilayer interactions and vulnerability to proteolytic enzymes, resulting in disorganization of the myelin sheath in the central nervous system. Therefore, citrullinated MBP (citMBP) has been suggested as a hallmark of demyelination, but how citMBP is implicated in prion diseases remains unknown. For the first time, we developed mouse monoclonal anti-citMBP IgG1 (clones 1B8, 1H1, and 3C6) and IgM (clone 3G5) antibodies that recognize human citMBP at its R25, R122, and R130 residues and at its C-terminal region (or the corresponding sites in mouse MBP), respectively. Using a biochemical, immunohistochemical, and immunogold-silver staining for electron microscopy techniques, we found that MBP residue R23 (corresponding to human R25) was specifically citrullinated, was stained as intense punctae in the corpus callosum, the striatum, and the cerebellar white matter, and was predominantly localized in disorganized myelin in the brains of scrapie-infected mice. In the brains of Creutzfeldt-Jakob disease (CJD) patients, MBP residues R25, R122, and R130 were markedly citrullinated and were stained as fibrils and punctae. In particular, white matter regions, such as the midbrain and the medulla, exhibited high levels of citMBP compared to other regions. However, the high levels of citMBP were not correlated with PAD2 expression. The clone 3G5 recognized significantly increased expression of the 18.5 kDa and/or 21.5 kDa variants of MBP in prion disease. Our findings suggest that significantly increased levels of citMBP may reflect demyelinating neuropathology, and that these newly developed antibodies may be useful for identifying demyelination.

Keywords

Myelin basic protein Citrullination Peptidylarginine deiminase Demyelination Neurodegeneration 

Notes

Acknowledgements

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2013R1A1A2009822 and NRF-2015R1D1A1A01059584) and by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (HI16C0965).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Byungki Jang
    • 1
  • Yong-Chul Jeon
    • 1
  • Hae-Young Shin
    • 1
  • Yun-Jung Lee
    • 1
  • Hyunji Kim
    • 1
  • Yoshitaka Kondo
    • 2
  • Akihito Ishigami
    • 2
  • Yong-Sun Kim
    • 1
    • 3
  • Eun-Kyoung Choi
    • 1
    • 4
  1. 1.Ilsong Institute of Life ScienceHallym UniversityAnyang, Gyeonggi-doRepublic of Korea
  2. 2.Molecular Regulation of AgingTokyo Metropolitan Institute of GerontologyTokyoJapan
  3. 3.Department of Microbiology, College of MedicineHallym UniversityChuncheon, Gangwon-doRepublic of Korea
  4. 4.Department of Biomedical GerontologyGraduate School of Hallym UniversityChuncheon, Gangwon-doRepublic of Korea

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