Deglycosylation was by the improved method.
Fetuin (glycoprotein from Sigma) has intrinsic microheterogeneity. Room temp works better than ice temperature. Most deglycosylation occurs in < 10min, but fetuin has resistant glycosyl residues which require more than 30 min to hydrolyze. Amount of fetuin should be reduced about 5fold in future experiments.
ATCV was inadvertently added to the deglycosylated ATCV sample.
Major capsid proteins of all phycodnaviruses are glycosylated, PBCV perhaps more than the others. Large virion proteins are also glycosylated. TFMSA deglycosylates cleanly w/o generating significant microheterogeneity. In fact capsid proteins appear much superior to fetuin (cleaner) as models for studying chemical deglycosylation.
Smaller proteins are missing from deglycosylated samples suggesting that precipitation requires more attention (ppt 30 min at -20C?).