Rationale: Influenza A viruses (IAVs) are still a threat to human health and life. The process of virus infection involves a series of biological regulations, such as signal transduction, that may be closely linked with the function of glycoproteins. However, the number and level of glycoproteins is low compared with other proteins in the whole protein pool. Methods: Viruses obtained from chicken embryos were purified by sucrose gradient centrifugation. PNGase F enzyme was then used to remove the glycan modification, followed by two-dimensional electrophoresis (2DE) to separate the hemagglutinin1 (HA1) glycoprotein. In-gel digestion was used to obtain peptides that were detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Results: Remarkably, we found five isoforms of HA1 with the same molecular weight but different isoelectric points. Furthermore, HA1 treatment with PNGase F enzyme changed all but one protein spot from 2DE, indicating that the different HA1 isoforms in 2DE were a result of different glycosylation modifications. Conclusions: The difference in isoelectric points of these HA1 isoforms was caused by glycan modification. This method provides a new approach for the study of glycosylation of the proteome for viruses or any other organisms.
|Number of pages||7|
|Journal||Rapid Communications in Mass Spectrometry|
|Publication status||Published - 30 Aug 2018|
Scopus Subject Areas
- Analytical Chemistry
- Organic Chemistry