Assay Methods for the Glycosyltransferases Involved in Synthesis of Bacterial Polysaccharides
Glycans play many important roles in bacterial biology and the complexity of the glycan structures requires biochemical assays in place to help characterize the biosynthetic pathways. Our focus has been on the use of enzymes from pathogens which make molecular mimics of host glycans. We have been examining glycosyltransferases that make strategic linkages in biologically active glycans which can be also exploited for potential therapeutic glycoconjugate synthesis. This chapter will provide details on assays for a variety of bacterial glycosyltransferases that we and others have used for the characterization of pathogen glycoconjugate biosynthetic pathways, and for the in vitro synthesis of human-like glycans produced by bacterial pathogens. The methods presented here should enable other assays to be developed for new pathway characterization.
Key wordsLipopolysaccharide Sialyltransferase BODIPY Enzyme assay Synthetic acceptor
We thank Dr. Hong-Ming Chen, University of British Columbia, for the gift of BODIPY-NHS. We thank Sussex Research Chemicals, for the gift of azido-sugars. The work was supported by grants to WW from GlycoNet doi: https://doi.org/10.13039/501100009056, and BJ was supported by an Erwin Schrödinger Fellowship.
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