2017年7月
WGA-based lectin affinity gel electrophoresis: A novel method for the detection of OGlcNAc-modified proteins
PLOS ONE
- ,
- ,
- 巻
- 12
- 号
- 7
- 開始ページ
- e0180714
- 終了ページ
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1371/journal.pone.0180714
- 出版者・発行元
- PUBLIC LIBRARY SCIENCE
Post-translational modification with O-linked beta-N-acetylglucosamine (O-GlcNAc) occurs selectively on serine and/or threonine residues of cytoplasmic and nuclear proteins, and dynamically regulates their molecular functions. Since conventional strategies to evaluate the O-GlcNAcylation level of a specific protein require time-consuming steps, the development of a rapid and easy method for the detection and quantification of an O-GlcNAcylated protein has been a challenging issue. Here, we describe a novel method in which O-GlcNAcylated and non-O-GlcNAcylated forms of proteins are separated by lectin affinity gel electrophoresis using wheat germ agglutinin (WGA), which primarily binds to N-acetylglucosamine residues. Electrophoresis of cell lysates through a gel containing copoly-merized WGA selectively induced retardation of the mobility of O-GlcNAcylated proteins, thereby allowing the simultaneous visualization of both the O-GlcNAcylated and the unmodified forms of proteins. This method is therefore useful for the quantitative detection of O-GlcNAcylated proteins.
- リンク情報
- ID情報
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- DOI : 10.1371/journal.pone.0180714
- ISSN : 1932-6203
- PubMed ID : 28686627
- Web of Science ID : WOS:000405464100095