Glycan microarrays for screening sialyltransferase

Authors: Ola Blixt Kirk Allin Ognian Bohorov Xiaofei Liu Hillevi AnderssonSand Julia Hoffmann Nahid Razi

Publish Date: 2007/10/04

Volume: 25, Issue: 1, Pages: 59-68

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Abstract

Here we demonstrate that glycan microarrays can be used for highthroughput acceptor specificity screening of various recombinant sialyltransferases Cytidine5′monophosphoNacetylneuraminic acid CMPNeu5Ac was biotinylated at position 9 of Nacetylneuraminic acid Neu5Ac by chemoenzymatic synthesis generating CMP9BiotNeu5Ac The activated sugar nucleotide was used as donor substrate for various mammalian sialyltranferases which transferred biotinylated sialic acids simultaneously onto glycan acceptors immobilized onto a microarray glass slide Biotinylated glycans detected with fluorescein–streptavidin conjugate to generate a specificity profile for each enzyme both confirming previously known specificities and reveal additional specificity information Human α26sialyltransferaseI hST6GalI also sialylates chitobiose structures GlcNAcβ14GlcNAcn including Nglycans rat α23sialyltransferase rST3GalIII tolerates fucosylated acceptors such as Lewisa human α23sialyltransferaseIV hST3GalIV broadly sialylates oligosaccharides of types 1–4 and porcine α23sialyltransferaseI pST3GalI sialylates gangliooligosaccharides and core 2 Oglycans in our array system Several of these sialyltransferases perform a substitution reaction and exchange a sialylated acceptor with a biotinylated sialic acid but are restricted to the most specific acceptor substrates Thus this method allows for a rapid generation of enzyme specificity information and can be used towards synthesis of new carbohydrate compounds and expand the glycan array compound library

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