O-linked sugars

Olivier Nosjean nosjean at univ-lyon1.fr
Thu Sep 26 05:23:30 EST 1996


In article (Dans l'article) <51rlt8$h22 at dux.dundee.ac.uk>,
(k.c.breen at dundee.ac.uk) wrote (écrivait) :

> I would like to check for the presence of Ser/Thr-O-GlcNAc on a protein.
> Looking through the appropiate catalogues, I cannot find a lectin that
> specifically labels this sugar group. I would be grateful if somebody could
> give me details of how I may assay for this particular sugar linkage.
> 
> Thanks,
> 
> Kieran Breen, Dept. of Pharmacology,                  
> University of Dundee,                   
> Ninewells Hospital & Medical School,    
> Dundee DD1 9SY,Scotland, U.K.                                              
>                
> k.c.breen at dundee.ac.uk                  

As to check for the presence of O-linked glycans, you can always try
enzymatic de-O-glycosylation (endo-alpha-N-acetylgalactosaminidase) and
check for glycans removal by SDS-PAGE. If it does not work, it does NOT
completely rule out the presence of Ser/Thr glycosylation. For instance,
substitution of the disaccharide bound to Ser/Thr will inhibit hydrolysis.
But if it works... you're in !

Chemical release is more efficient (beta elimination), but requires more
material.

Many practical details on how to handle this kind of question in Methods
on glycoconjugates (1995) Edited by Andre Verbert, hardwood academic
publishers, Switzerland.


...Oli

-----
Olivier Nosjean
Laboratory of Biological Physico-chemistry
Universite C. Bernard-Lyon 1, France

nosjean at univ-lyon1.fr
-----


_  _  _
 \/ \/
  {O}        Olivier & Marie-Caroline
  |_|            F650 is great !
  |||              
   |          nosjean at univ-lyon1.fr



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