WGA-based lectin affinity gel electrophoresis: A novel method for the detection of O-GlcNAc-modified proteins.
Post-translational modification with O-linked β-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 spe...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2017-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC5501588?pdf=render |
| _version_ | 1818899884515786752 |
|---|---|
| author | Yuji Kubota Ko Fujioka Mutsuhiro Takekawa |
| author_facet | Yuji Kubota Ko Fujioka Mutsuhiro Takekawa |
| author_sort | Yuji Kubota |
| collection | DOAJ |
| description | Post-translational modification with O-linked β-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 copolymerized 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. |
| first_indexed | 2024-12-19T19:55:03Z |
| format | Article |
| id | doaj.art-592ce765d5764e06b7e21664a4703941 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-19T19:55:03Z |
| publishDate | 2017-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-592ce765d5764e06b7e21664a47039412022-12-21T20:07:50ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01127e018071410.1371/journal.pone.0180714WGA-based lectin affinity gel electrophoresis: A novel method for the detection of O-GlcNAc-modified proteins.Yuji KubotaKo FujiokaMutsuhiro TakekawaPost-translational modification with O-linked β-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 copolymerized 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.http://europepmc.org/articles/PMC5501588?pdf=render |
| spellingShingle | Yuji Kubota Ko Fujioka Mutsuhiro Takekawa WGA-based lectin affinity gel electrophoresis: A novel method for the detection of O-GlcNAc-modified proteins. PLoS ONE |
| title | WGA-based lectin affinity gel electrophoresis: A novel method for the detection of O-GlcNAc-modified proteins. |
| title_full | WGA-based lectin affinity gel electrophoresis: A novel method for the detection of O-GlcNAc-modified proteins. |
| title_fullStr | WGA-based lectin affinity gel electrophoresis: A novel method for the detection of O-GlcNAc-modified proteins. |
| title_full_unstemmed | WGA-based lectin affinity gel electrophoresis: A novel method for the detection of O-GlcNAc-modified proteins. |
| title_short | WGA-based lectin affinity gel electrophoresis: A novel method for the detection of O-GlcNAc-modified proteins. |
| title_sort | wga based lectin affinity gel electrophoresis a novel method for the detection of o glcnac modified proteins |
| url | http://europepmc.org/articles/PMC5501588?pdf=render |
| work_keys_str_mv | AT yujikubota wgabasedlectinaffinitygelelectrophoresisanovelmethodforthedetectionofoglcnacmodifiedproteins AT kofujioka wgabasedlectinaffinitygelelectrophoresisanovelmethodforthedetectionofoglcnacmodifiedproteins AT mutsuhirotakekawa wgabasedlectinaffinitygelelectrophoresisanovelmethodforthedetectionofoglcnacmodifiedproteins |