Phos-tag SDS-PAGE analysis of complicated phosphorylation of a neurodegenerative disease protein tau

• Kimura Taeko

  Department of Functional Brain Imaging Research, National Institute for Quantum and Radiological Science and Technology Laboratory of Molecular Neuroscience, Department of Biological Sciences, Tokyo Metropolitan University

• Hisanaga Shinichi

  Laboratory of Molecular Neuroscience, Department of Biological Sciences, Tokyo Metropolitan University

Bibliographic Information

Other Title

• 複雑怪奇なリン酸化タンパク質タウのPhos-tag解析

Abstract

Tau is a microtubule-associated protein primarily expressed in the axons of neurons. Tau regulates microtubule dynamics and transport of organelles along microtubules in axons. These functions of tau are regulated by phosphorylation with a number of protein kinases. Tau is also a major component of neurofibrillary tangles (NFTs) in the brains of patients with tauopathies, including Alzheimer’s disease (AD), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). Tau in NFTs is hyperphosphorylated, and therefore, the phosphorylation of tau has been intensively investigated. To date, more than 40 phosphorylation sites have been identified in aggregated tau. However, the physiological and pathological roles of the respective phosphorylation sites and how their phosphorylation is regulated are unclear. This could be due to the lack of the proper method of analyzing tau phosphorylation. Most of previous phosphorylation studies were conducted with a repertory of phosphorylation-site specific antibodies. While they provide relative changes of respective phosphorylation sites, it is hard to estimate total or absolute phosphorylation. To overcome these problems, we have recently applied the Phos-tag technique to the analysis of tau phosphorylation. This method separated tau into many bands on SDS-PAGE depending on phosphorylation, making it possible to analyze tau phosphorylation quantitatively and combinatorially. We identified phosphor-isotypes of tau expressed in cultured cells and found the abundance of nonphosphorylated tau in mouse and human tauopathy brains. Our results demonstrate that the Phos-tag would be a powerful method to characterize in vivo physiological and pathological phosphorylation of tau.

Journal

• Electrophoresis Letters

  Electrophoresis Letters 61 (2), 49-52, 2017

  Japanese Electrophoresis Society

Keywords

• Alzheimer’s disease
• tauopathy
• tau
• phosphoryaltion
• Phos-tag SDS-PAGE

Details

• CRID: 1390001205763372288
• NII Article ID: 130006207357
• DOI: 10.2198/electroph.61.49
• ISSN: 21892636
• ISSN: 21892628
• Web Site: https://www.jstage.jst.go.jp/article/electroph/61/2/61_49/_pdf
• Web Site: https://search.jamas.or.jp/link/ui/2018148620
• Text Lang: ja
• Data Source:
  • JaLC
  • Crossref
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed