| Status | 已發表Published |
| Quantitative characterization of Site-Specific glycosylation of glycoproteins - Using Human transferrin as a model | |
Wei, R. ; Poon, C. W.
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| 2017-06-21 | |
| Source Publication | 4th Macau Symposium on Biomedical Sciences 2017
 |
| Abstract | Background: Glycoproteins carrying aberrant glycosylations, such as carbohydrate deficient transferrin, have been used as disease biomarkers. Various assays have been developed to quantify glycosylation changes of glycoprotein biomarkers, but none of them can reliably quantify glycosylation patterns at specific glycosylation sites. Deciphering detailed information of site-specific glycosylation not only can help us understanding the structural and functional relationship of glycoproteins, but also can create new opportunities in disease diagnostics. Objective and Method: Use human transferrin as a glycoprotein model, we aimed to identify experimental factors that affect mass spectrometry-based quantitative characterization of carbohydrate sides at the individual glycosylation sites. Results and Discussion: We observed that trypsin digestion protocols significantly affected the relative signal intensities of individual glycosylated peptides released from transferrin. Furthermore, our data showed that trypsin digestion resulted in insufficient release of the tryptic peptide containing the Asn 413 glycosylation site when compared to the one containing the Asn611 glycosylation site, subsequently resulting in lower signal intensities of both glycosylated and non-glycosylated forms of the former tryptic peptide. Introduction of an additional protein digestion step using LysC could alleviate such digestion bias, suggesting that higher miscleavage rate at lysine residues was the major cause of the digestion bias. Conclusion: We have illustrated that trypsin digestion alone may not be able to release glycopeptides containing different glycosylation sites in an unbiased manner. Carefully optimization of the digestion protocol and inclusion of an additional endopeptidase for protein digestion can help alleviating the quantification bias due to digestion bias. |
| Keyword | glycoprotein proteomics glycosylation mass spectrometry |
| Language | 英語English |
| The Source to Article | PB_Publication |
| PUB ID | 33382 |
| Document Type | Conference paper |
| Collection | DEPARTMENT OF BIOMEDICAL SCIENCES |
| Corresponding Author | Poon, C. W. |
| Recommended Citation GB/T 7714 | Wei, R.,Poon, C. W.. Quantitative characterization of Site-Specific glycosylation of glycoproteins - Using Human transferrin as a model[C], 2017. |
| APA | Wei, R.., & Poon, C. W. (2017). Quantitative characterization of Site-Specific glycosylation of glycoproteins - Using Human transferrin as a model. 4th Macau Symposium on Biomedical Sciences 2017. |
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