Florescent substrates for glycosyltransferases?

Wed Sep 13 11:44:08 EST 1995

 Canadian group of Prof. Ole Hidsgaul has recently reported the 
 use of tetramethylrhodamine as the fluorescent label on 
 oligosaccharides for the assay of both glycosidases and 
 glycosyltransferases.  This is probably the most sensitive 
 method reported todate.  Detection of 100 molecules of enzyme 
 substrate product can be achieved by using capillary 
 electrophoresis with laser-induced fluorescence detection.
 Following are the references:
 Glycobiology, 4 (1994) 239-242
 Anal. Biochem., 227 (1995) 368-376
 Anal. Biochem., 205 (1992) 108-114 (Label: ANDA)
 J. Chromatography B, 657 (1994) 307-313
 Complex Carbohydrates in Drug Research (Alferd Benzon Symposium 
 36), K. Bock and H. Clausen, Eds., Munksgaard Intl. Publ., 
 Copenhagen, (1994) 104-113.
 In addition to the above labels, Pyridylamino-oligosaccharides 
 can also be used as substrates as suggested by Ian Wilson.
 Shaheer H. Khan
 Applied Biosystems
 Foster City, CA
 Disclaimer: Only and Only my opinion, my employer has nothing 
 to do with it.
 Dear Colleagues,
 I have noticed that one can use commercially available 
 AMC-labelled oligosaccharides as substrates 
 for glycosidase assays.
 My question is whether there is any experience around with 
 using similarly labeelled  
 oligosaccharides as substrates for glycosyltransferases. If yes 
 then how far the label should 
 be from the reaction centre and what might be the optimal 
 method for following the reaction, e.g. 
 TLC or HPLC.I am mostly thinking about assaying the activity  
 of  alpha- galactosyl transferase.
 Armin Sepp, PhD
 Armin.Sepp at bbsrc.ac.uk

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