ISSN:
0947-3440
Keywords:
Sialyltransferase
;
Acceptor-specific assay
;
LacNAc
;
Fluorescein labelling
;
Chemistry
;
Organic Chemistry
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Chemistry and Pharmacology
Notes:
A novel HPLC assay system for sialyltransferase activity based on the use of fluorescein-labelled acceptor oligosaccharides is described. The fluorescein-labelled disaccharides βGal-(1→4)-βGlc-OR (4), βGal-(1→4)-βGlcNAc-OR (17), and βGal-(1→3)-βGlcNAc-OR (22) where OR consists of a six carbon spacer with fluorescein attached, were synthesised. Synthetic standard products were produced chemo-enzymatically on a preparative scale to yield fluorescein-labelled trisaccharides. The use of reverse phase HPLC with an ionpairing agent allowed the separation of starting materials from products and separation of the two isomeric trisaccharides αNeu5Ac-(2→3/6)-βGal-(1→4)-βGlcNAc-OR (24 and 25), so that the assay could be used to measure the different silalyltransferase activities in a mixture. The assay was successfully applied to the detection of sialyltransferase activity of commercially available enzymes and a crude preparation of bovine colostrum. The predominant sialyltransferase activity in bovine colostrum adds sialic acid α(2-6) to 17.
Additional Material:
2 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/jlac.199619961111
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