AI Article Synopsis
- A new method called "Glyco-seek" allows for the non-destructive analysis of O-GlcNAcylated proteins using a combination of labeling, proximity ligation, and quantitative PCR.
- This technique offers extremely high sensitivity, enabling the detection of very low amounts (attomoles) of glycoproteins in cellular samples.
- Glyco-seek can operate alongside traditional assays to measure both O-GlcNAcylation levels and total protein amounts, providing a comprehensive view of protein modifications without the need for complex isolation processes.
Article Abstract
We report a non-destructive biochemical technique, termed "Glyco-seek", for analysis of O-GlcNAcylated proteins. Glyco-seek combines chemoenzymatic labeling, proximity ligation, and quantitative polymerase chain reaction to detect O-GlcNAcylated proteins with ultrahigh sensitivity. Our glycan-specific assay can be paired with traditional proximity ligation assays to simultaneously determine the change in total protein levels. We show that Glyco-seek detects attomoles of glycoproteins of interest from cell lysates, with sensitivity several orders of magnitude higher than that of current techniques. We used the method to directly assay the O-GlcNAcylation status of a low-abundance transcription factor from cell lysates without need for isolation or enrichment.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5327792 | PMC |
| http://dx.doi.org/10.1021/jacs.6b03861 | DOI Listing |
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