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| Content Provider | PubMed Central |
|---|---|
| Author | Zhou, Hui Froehlich, John W. Briscoe, Andrew C. Lee, Richard S. |
| Copyright Year | 2013 |
| Abstract | Current strategies to study N-glycoproteins in complex samples are often discrete, focusing on either N-glycans or N-glycosites enriched by sugar-based techniques. In this study we report a simple and rapid sample preparation platform, the GlycoFilter, which allows a comprehensive characterization of N-glycans, N-glycosites, and proteins in a single workflow. Both PNGase F catalyzed de-N-glycosylation and trypsin digestions are accelerated by microwave irradiation and performed sequentially in a single spin filter. Both N-glycans and peptides (including de-N-glycosylated peptides) are separately collected by filtration. The condition to effectively collect complex and heterogeneous N-glycans was established on model glycoproteins, bovine ribonuclease B, bovine fetuin, and human serum IgG. With this platform, the N-glycome, N-glycoproteome and proteome of human urine and plasma were characterized. Overall, a total of 865 and 295 N-glycosites were identified from three pairs of urine and plasma samples, respectively. Many sites were defined unambiguously as partially occupied by the detection of their nonsugar-modified peptides (128 from urine and 61 from plasma), demonstrating that partial occupancy of N-glycosylation occurs frequently. Given the likely high prevalence and variability of partial occupancy, glycoprotein quantification based exclusively on deglycosylated peptides may lead to inaccurate quantification. |
| Related Links | http://dx.doi.org/10.1074/mcp.m113.027953 |
| Ending Page | 2991 |
| Page Count | 11 |
| Starting Page | 2981 |
| File Format | |
| ISSN | 15359476 |
| e-ISSN | 15359484 |
| Journal | Molecular & Cellular Proteomics : MCP |
| Issue Number | 10 |
| Volume Number | 12 |
| Language | English |
| Publisher | The American Society for Biochemistry and Molecular Biology |
| Publisher Date | 2013-10-01 |
| Access Restriction | Open |
| Rights Holder | The American Society for Biochemistry and Molecular Biology |
| Subject Keyword | Analytical Chemistry Biochemistry Molecular Biology Research in Higher Education |
| Content Type | Text |
| Resource Type | Article |
| Subject | Medicine Analytical Chemistry Molecular Biology Biochemistry |
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