Study of stability of glycosylated and deglycosylated PA83 variants using SDS-PAGE analysis.

<p>Plant produced, glycosylated PA83, and <i>in vivo</i> Endo H or PNGase F deglycosylated forms of PA83 were purified as described in Materials and Methods. (A) The purified plant produced PA83 variants were stored for 1 hour at 37°C or for 72 hours at 4°C and analyzed by SDS-PAGE. Lanes were loaded with ~8.0 μg per lane for glycosylated, Endo H or PNGase F <i>in vivo</i> deglycosylated plant produced PA83 proteins. (B) Purified, plant produced, glycosylated PA83, and <i>in vivo</i> deglycosylated (co-expressed with Endo H or PNGase F) and <i>in vitro</i> deglycosylated (by commercial Endo H) proteins were incubated at 37°C for 1, 4, 8, 16 and 24 hours, and analyzed in SDS-PAGE. Lanes were loaded with ~5.0 μg per lane for each sample. M- color prestained protein standard (New England Biolabs).