New England Biolabs, 240 County Road, Ipswich, MA, 01938, USA.
Department of Parasitology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA, Leiden, The Netherlands.
Sci Rep. 2018 Jun 22;8(1):9504. doi: 10.1038/s41598-018-27797-0.
Exoglycosidases are often used for detailed characterization of glycan structures. Bovine kidney α-fucosidase is commonly used to determine the presence of core α1-6 fucose on N-glycans, an important modification of glycoproteins. Recently, several studies have reported that removal of core α1-6-linked fucose from N-glycans labeled with the reactive N-hydroxysuccinimide carbamate fluorescent labels 6-aminoquinolyl-N-hydroxysuccinimidylcarbamate (AQC) and RapiFluor-MS is severely impeded. We report here the cloning, expression and biochemical characterization of an α-fucosidase from Omnitrophica bacterium (termed fucosidase O). We show that fucosidase O can efficiently remove α1-6- and α1-3-linked core fucose from N-glycans. Additionally, we demonstrate that fucosidase O is able to efficiently hydrolyze core α1-6-linked fucose from N-glycans labeled with any of the existing NHS-carbamate activated fluorescent dyes.
外切糖苷酶常用于详细表征聚糖结构。牛肾α-岩藻糖苷酶常用于确定 N-聚糖上核心α1-6 岩藻糖的存在，这是糖蛋白的一个重要修饰。最近，有几项研究报告称，从用反应性 N-羟基琥珀酰亚胺碳酸酯荧光标签 6-氨基喹啉基-N-羟基琥珀酰亚胺碳酸酯（AQC）和 Rapifluor-MS 标记的 N-聚糖中去除核心α1-6 连接的岩藻糖会受到严重阻碍。我们在此报道了一种来自 Omnitrophica 细菌的α-岩藻糖苷酶（称为岩藻糖苷酶 O）的克隆、表达和生化特性。我们表明，岩藻糖苷酶 O 可以有效地从 N-聚糖中去除α1-6-和α1-3-连接的核心岩藻糖。此外，我们证明岩藻糖苷酶 O 能够有效地从用任何现有的 NHS-碳酸酯活化荧光染料标记的 N-聚糖中水解核心α1-6 连接的岩藻糖。
