Open AccessExperimental Design Considerations for In Vitro Non‐Natural Glycan Display via Metabolic Oligosaccharide Engineering
Author(s) -
Tan Elaine,
Almaraz Ruben T.,
Khanna Hargun S.,
Du Jian,
Yarema Kevin J.
Publication year - 2010
Publication title -
current protocols in chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.503
H-Index - 14
ISSN - 2160-4762
DOI - 10.1002/9780470559277.ch100059
Subject(s) - glycan , oligosaccharide , glycosylation , metabolic pathway , computational biology , metabolic engineering , biochemistry , cell , chemistry , biology , microbiology and biotechnology , metabolism , enzyme , glycoprotein
Metabolic oligosaccharide engineering (MOE) refers to a technique where non‐natural monosaccharide analogs are introduced into living biological systems. Once inside a cell, these compounds intercept a targeted biosynthetic glycosylation pathway and in turn are metabolically incorporated into cell‐surface‐displayed oligosaccharides where they can modulate a host of biological activities or be exploited as “tags” for bio‐orthogonal and chemoselective ligation reactions. Undertaking a MOE experiment can be a daunting task based on the growing repertoire of analogs now available and the ever increasing number of metabolic pathways that can be targeted; therefore, a major emphasis of this article is to describe a general approach for analog design and selection and then provide protocols to ensure safe and efficacious analog usage by cells. Once cell‐surface glycans have been successfully remodeled by MOE methodology, the stage is set for probing changes to the myriad cellular responses modulated by these versatile molecules. Curr. Protoc. Chem. Biol . 2:171‐194 © 2010 by John Wiley & Sons, Inc.