exoglycosidase activity on native glycoproteins

Dr. David Hoover hooverd at CRYSV1.NCIFCRF.GOV
Tue Jun 17 15:00:22 EST 1997

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.


Dr. David Hoover, PhD
Macromolecular Laboratory Group
P.O. Box B, Bldg. 539
Frederick, MD 21702-1201
Phone: (301)-846-5031
FAX: (301)-846-7101

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