Method for analyzing protein O-glycosylation sites
A technology for glycosylation sites and proteins, which is applied in the field of analyzing protein O-glycosylation sites, can solve the problems of undiscovered, difficult to control chemical methods, and difficult to analyze O-glycosylation sites, and achieves a simple method. Effect
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[0007] A method for analyzing protein O-glycosylation sites, the steps are as follows:
[0008] (1) After the proteome sample or a single protein sample is cleaved with endonuclease, the N-glycosidase and exoglycosidase are combined to cut off the N-sugar chain and part of the O-sugar chain, and dried under reduced pressure to obtain the Omics protein samples or single protein samples of GalNAc sugar-tagged peptides and non-glycopeptides;
[0009] (2) Load the omics protein samples with GalNAc sugar tags and non-glycopeptides prepared in step (1) on the durian lectin column for separation, and collect the enzyme-digested peptides with GalNAc sugar tags The peptide solution is dried under reduced pressure to prepare the hismic protein sample;
[0010] (3) Use the single protein sample prepared in step (1) or the hismic protein sample prepared in step (2) to 18 Separation by reversed-phase chromatographic column, and then detection by high-resolution mass spectrometer in posit...
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