Tools and tactics to define specificity of metabolic chemical reporters

Author:

Mukherjee, Mana Mohan

Bond, Michelle R

Abramowitz, Lara K

Biesbrock, Devin

Woodroofe, Carolyn C

Kim, Eun Ju

Swenson, Rolf E

Hanover, John A
Author Address

Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, United States., Frederick National Laboratory for Cancer Research, National Cancer Institute, Fredrick, MD, United States., Department of Chemistry Education, Daegu University, Gyeongsan-si, South Korea.,

Abstract:

Metabolic chemical reporters (MCRs) provide easily accessible means to study glycans in their native environments. However, because monosaccharide precursors are shared by many glycosylation pathways, selective incorporation has been difficult to attain. Here, a strategy for defining the selectivity and enzymatic incorporation of an MCR is presented. Performing ß-elimination to interrogate O-linked sugars and using commercially available glycosidases and glycosyltransferase inhibitors, we probed the specificity of widely used azide (Ac4GalNAz) and alkyne (Ac4GalNAlk and Ac4GlcNAlk) sugar derivatives. Following the outlined strategy, we provide a semiquantitative assessment of the specific and non-specific incorporation of this bioorthogonal sugar (Ac4GalNAz) into numerous N- and O-linked glycosylation pathways. This approach should be generally applicable to other MCRs to define the extent of incorporation into the various glycan species. Copyright © 2023 Mukherjee, Bond, Abramowitz, Biesbrock, Woodroofe, Kim, Swenson and Hanover.

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