The Use of Two-Dimensional SDS-PAGE to Analyze the Glycan Heterogeneity of the Respiratory Syncytial Virus Fusion Protein
The respiratory syncytial virus (RSV) fusion (F) protein is synthesized as an inactive precursor (F0), which subsequently undergoes post-translational cleavage to give the disulphide bond-linked F1 and F2 subunits. The methodology detailing the use of two-dimensional electrophoresis, endoglycosidases, and α-mannosidase inhibitors, as applied to investigating F protein glycan maturation, is given. Examples are used to show how this methodology was used to provide evidence for glycan heterogeneity within the mature F protein.
Key WordsProteomics F protein respiratory syncytial virus deoxymannojirimycin Swainsonine
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