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

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Selective Oxidation of N-Glycolylneuraminic Acid Using an Engineered Galactose Oxidase Variant
ACS Catalysis ( IF 11.3 ) Pub Date : 2019-08-05 00:00:00 , DOI: 10.1021/acscatal.9b02873
Ashley P. Mattey ₁ , William R. Birmingham ₁ , Peter Both ₁ , Nico Kress ₁ , Kun Huang ₁ , Jolanda M. van Munster ₁ , Gregory S. Bulmer ₁ , Fabio Parmeggiani ₁ , Josef Voglmeir ₂ , Juana E. R. Martinez ₃ , Nicholas J. Turner ₁ , Sabine L. Flitsch ₁

Affiliation

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.

中文翻译：

使用工程化的半乳糖氧化酶变体选择性氧化N-乙二醇神经氨酸

N-甘氨酰神经氨酸（Neu5Gc）是动物中常见的细胞表面配体，不能在人体内生物合成，但可以在人体组织中从饮食来源（例如红肉）中获得。理解对人类健康这个潜在的免疫原性聚糖的相关性是很重要的，并且需要选择性探测方法，可以从它的生物合成前体区分的Neu5Gc，在人类是共同（即，Ñ-乙酰神经氨酸（Neu5Ac））。在这里，我们证明Neu5Gc可以被半乳糖氧化酶的工程变体选择性氧化，而不会与Neu5Ac发生任何反应。Neu5Gc本身的氧化可以对醛产物进行全光谱表征。此外，我们显示，Neu5Gc是典型动物寡糖基序的一部分，并且与蛋白质连接的N-聚糖连接时，也会被氧化。Neu5Gc的氧化引入了可以正交标记的生物正交功能。我们证明与唾液酸酶介导的水解结合，此两种酶系统可以为选择性检测复杂生物样品（例如生物药物α酸糖蛋白）中的Neu5Gc提供有用的工具。

更新日期：2019-08-05

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