Combinatorial Chemoenzymatic Synthesis and High-throughput Screening of Sialosides

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Title Combinatorial Chemoenzymatic Synthesis and High-throughput Screening of Sialosides
Author Chokhawala, Harshal A.; Huang, Shengshu; Lau, Kam; Yu, Hai; Cheng, Jiansong; Thon, Vireak; Hurtado-Ziola, Nancy; Guerrero, Juan A.; Varki, Ajit; Chen, Xi
Journal Name ACS Chemical Biology
Year Published 2008
Place of publication United States
Publisher American Chemical Society
Abstract Although the vital roles of structures containing sialic acid in biomolecular recognition are well documented, limited information is available on how sialic acid structural modifications, sialyl linkages, and the underlying glycan structures affect the binding or the activity of sialic acid-recognizing proteins and related downstream biological processes. A novel combinatorial chemoenzymatic method has been developed for the highly efficient synthesis of biotinylated sialosides containing different sialic acid structures and different underlying glycans in 96-well plates from biotinylated sialyltransferase acceptors and sialic acid precursors. By transferring the reaction mixtures to NeutrAvidin-coated plates and assaying for the yields of enzymatic reactions using lectins recognizing sialyltransferase acceptors but not the sialylated products, the biotinylated sialoside products can be directly used, without purification, for high-throughput screening to quickly identify the ligand specificity of sialic acid-binding proteins. For a proof-of-principle experiment, 72 biotinylated α2,6-linked sialosides were synthesized in 96-well plates from 4 biotinylated sialyltransferase acceptors and 18 sialic acid precursors using a one-pot three-enzyme system. High-throughput screening assays performed in NeutrAvidin-coated microtiter plates show that whereas Sambucus nigra Lectin binds to α2,6-linked sialosides with high promiscuity, human Siglec-2 (CD22) is highly selective for a number of sialic acid structures and the underlying glycans in its sialoside ligands.
Peer Reviewed Yes
Published Yes
Alternative URI http://dx.doi.org/10.1021/cb800127n
Copyright Statement Self-archiving of the author-manuscript version is not yet supported by this journal. Please refer to the journal link for access to the definitive, published version or contact the authors for more information.
Volume 3
Issue Number 9
Page from 567
Page to 576
ISSN 1554-8937
Date Accessioned 2011-03-16
Date Available 2011-05-03T04:47:23Z
Language en_AU
Faculty Faculty of Science, Environment, Engineering and Technology
Subject Organic Chemical Synthesis
URI http://hdl.handle.net/10072/38543
Publication Type Journal Articles (Refereed Article)
Publication Type Code c1x

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