at11 at ukc.ac.uk (A.Thomson) wrote:
>I am looking for a method to separate a population of N-linked and O-linked
>Glycoproteins.
>One of the proteins has both forms of glycosylation present with the other
>having only O-linked glycosylation
>I have considered some form of affinity chromatography using Con A agarose
>but i am unsure as to whether this will be specific only for N-linked
>oligosaccharides.
>>A. Thomson.
Dear A. Thomson
Although I'm not exactly an Con A expert, I now the following for sure:
BIANTENNARY-type glycopeptides are bound and eluted with a low hapten
concentration (10 mM alpha-methylglucoside),
while OLIGOMANNOSE/HYBRID-type glycopeptides are bound up to an high hapten
concentration (100 mM alpha-methylmannoside).(Ref. Methods in Enzymology vol.
230, pp. 66-87)
This means that N-linked glycoproteins with tri- and tetraantennary
oligosaccharides are not retarded, nor bound, to a Con A column, which makes
your separation of N-linked and O-linked rather tricky.
Corne Stroop (cjms at boc.chem.ruu.nl)
Bijvoet Center for Biomolecular Research
Department of Bio-organic Chemistry
Utrecht University
Padualaan 8, NL-3584 CH
Utrecht, the Netherlands
