O-linked glycosylation

Bob Lauder R.Lauder at lancaster.ac.uk
Mon Mar 20 08:22:32 EST 1995


In article <3kc99i$nvt at mserv1.dl.ac.uk>, "Tim Dudgeon" <dudgeon at britbio.co.uk> says:
>
>I have ..partially glycosylated with O-liked sugar (there are no N-linked sites in the
>protein). I wish to seperate the glycosylated form the unglycosylated protein.

>I have tried using ConA-Sepharose (Pharmacia) 

Con-A will almost certainly not bind your O-linked glycan chains. Con-A does have a use
for the separation of O-linked from N-linked chains, which it will, in general, bind.

>and Glycine-Max (Sigma) without

I have no experience of this lectin, anybody else know about it?

>I wish to seperate the glycosylated form the unglycosylated protein.

You could try a different approach - if the glycan chains are charged (sulphated ?) 
then an ion exchange column may be useful. Are the chains capped by Sialic acid ? 
if so then Sambucus nigra will bind (2-6)-linked Sialic acids. The Lectin for 
(2-3)-linked Sialic acid, Maccia, has been less sucessful in my hands (columns?).

Hope this helps a little.

================================================================
Bob Lauder				R.Lauder at lancaster.ac.uk
Division of Biological Sciences		Tel +(44)(0)1524 65201
Lancaster University, Lancaster, UK	Fax +(44)(0)1524 843854

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