Selective Oxidation of N-Glycolylneuraminic Acid Using an Engineered Galactose Oxidase Variant

Ashley P Mattey, William Birmingham, Peter Both, Nico Kress, Kun Huang, JM van Munster, Gregory S Bulmer, Fabio Parmeggiani, Josef Voglmeir, Juana Martinez, Nicholas Turner, Sabine L Flitsch*

*Corresponding author for this work

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

N-Glycolylneuraminic acid (Neu5Gc) is a common cell-surface ligand in animals which is not biosynthesized in humans, but it can be acquired in human tissue from dietary sources such as red meat. It is important to understand the relevance of this potentially immunogenic glycan on human health, and selective detection methods are needed that can distinguish Neu5Gc from its biosynthetic precursor that is common in humans (i.e., N-acetylneuraminic acid (Neu5Ac)). Here, we demonstrate that Neu5Gc can be selectively oxidized by an engineered variant of galactose oxidase without any reaction toward Neu5Ac. Oxidation of Neu5Gc itself allowed for the full spectroscopic characterization of the aldehyde product. In addition, we show that Neu5Gc is also oxidized when it is part of a typical animal oligosaccharide motif and when it is attached to a protein-linked N-glycan. Oxidation of Neu5Gc introduces bioorthogonal functionality that can be exclusively labeled. We demonstrate that in combination with sialidase-mediated hydrolysis, this two-enzyme system can provide a useful tool for the selective detection of Neu5Gc in complex biological samples such as the biopharmaceutical alpha acid glycoprotein.
Original languageEnglish
Pages (from-to)8208–8212
JournalACS Catalysis
Volume9
Issue number9
Early online date5 Aug 2019
DOIs
Publication statusPrint publication - 6 Sep 2019
Externally publishedYes

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    Mattey, A. P., Birmingham, W., Both, P., Kress, N., Huang, K., van Munster, JM., Bulmer, G. S., Parmeggiani, F., Voglmeir, J., Martinez, J., Turner, N., & Flitsch, S. L. (2019). Selective Oxidation of N-Glycolylneuraminic Acid Using an Engineered Galactose Oxidase Variant. ACS Catalysis, 9(9), 8208–8212. https://doi.org/10.1021/acscatal.9b02873