Does anyone have experiences with exoglycosidase reactions on native, non-
denatured glycoproteins? I have a glycoprotein with complex N-linked
oligosaccarides which, upon removal of the sugars by PNGase F, becomes
irreversibly denatured. Other endoglycosidases have no effect, due to the
complexity of the oligosaccarides. I would like to remove the terminal
sialic acids, galactoses, and glcNAcs.
Are there exoglycosidases available with high enough activity to allow this?
I understand there are better ways around this problem (expression in other,
more restrictive glycosylating systems), but for various reasons these options
are closed. A discussion of the quality of available exoglycosidases (such
as Sigma, Boehringer Mannheim, NEB, Glyko, etc.) would be very helpful.
Thanks!
-------------------------------------------------------------
Dr. David Hoover, PhD
Macromolecular Laboratory Group
ABL/NCI-FCRDC
P.O. Box B, Bldg. 539
Frederick, MD 21702-1201
Phone: (301)-846-5031
FAX: (301)-846-7101
