Lectin binding profiles on sucrose purified VLP. LASV Z+GPC+NP VLP fractions obtained from sucrose gradient sedimentation corresponding to those in Figure 1A were subjected to SDS-PAGE (3A) and lectin binding analysis on proteins transferred to nitrocellulose membranes (3B). A combination of agglutinins, GNA (Galanthus nivalis), SNA (Sambucus nigra), MAA (Maackia amurensis), PNA (Peanut), and DSA (Datura stramonium), were combined and used to probe VLP fractions 1 through 9 (3B, lanes 1 - 9). LASV NP, GP1, and GP2 generated in E. coli were used as unglycosylated protein controls (3B, lane 10). A combination of four glycoproteins was used as positive controls for lectin binding: carboxypeptidase Y (63 kDa), transferrin (80 kDa), fetuin (68, 65, 61 kDa), and asialofetuin (61, 55, 48 kDa) (3B, lane 11). For visual comparison purposes an SDS-PAGE gel was run with the same VLP fractions, stained with Coomassie BB-R250, and photographed (3A, lanes 1 - 9). LASV Z, Z+GPC+NP, Z+GPC, Z+NP VLP purified through 20% sucrose cushions were similarly analyzed for glycan binding (3C, lanes 1 - 4, respectively). The relative positions of GPC, GP1, and GP2 are noted to the left of the gel. Protein molecular weights in kDa are noted to the right of each image.
Branco et al. Virology Journal 2010 7:279 doi:10.1186/1743-422X-7-279