| RRC ID |
52303
|
| Author |
Kubota Y, Fujioka K, Takekawa M.
|
| Title |
WGA-based lectin affinity gel electrophoresis: A novel method for the detection of O-GlcNAc-modified proteins.
|
| Journal |
PLoS One
|
| Abstract |
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.
|
| Volume |
12(7)
|
| Pages |
e0180714
|
| Published |
2017-7-7
|
| DOI |
10.1371/journal.pone.0180714
|
| PII |
PONE-D-16-35933
|
| PMID |
28686627
|
| PMC |
PMC5501588
|
| MeSH |
Acetylglucosamine / chemistry*
Acetylglucosamine / metabolism
Electrophoresis
Glycosylation
Protein Processing, Post-Translational*
Proteins / chemistry
Proteins / isolation & purification
Proteins / metabolism*
Wheat Germ Agglutinins / chemistry
|
| IF |
2.74
|
| Times Cited |
4
|
| Resource |
| Human and Animal Cells |
293(RCB1637)
COS-7(RCB0539) |
