NIBRT GlycoScience Group, National Institute for Bioprocessing, Research and Training , Foster's Avenue, Mount Merrion, Blackrock, Co., Dublin, Ireland.
Department of Chemistry, Maynooth University , Maynooth, Kildare Ireland.
J Proteome Res. 2017 Nov 3;16(11):4237-4243. doi: 10.1021/acs.jproteome.7b00580.
Here we report evidence that new aminoquinoline N-glycan fluorescent labels interfere with the release of core α(1-6) fucose from N-glycans by bovine kidney α-l-fucosidase (BKF). BKF is a commonly employed exoglycosidase for core α(1-6) fucose determination. Molecular simulations of the bound and unbound Fuc-α(1-6)-GlcNAc, where GlcNAc is situated at the reducing end for all N-glycans, suggest that the reduced BKF activity may be due to a nonoptimal fit of the highest populated conformers in the BKF active binding site at room temperature. Population analysis and free energy estimates suggest that an enhanced flexibility of the labeled sugar, which facilitates recognition and binding, can be achievable with extended reaction conditions. We provide these experimental conditions using a sequential exoglycosidase digestion array using high concentrations of BKF.
在这里，我们报告了新的氨基喹啉 N-聚糖荧光标记物干扰牛肾α-L-岩藻糖苷酶（BKF）从 N-聚糖释放核心α（1-6）岩藻糖的证据。BKF 是一种常用于核心α（1-6）岩藻糖测定的外切糖苷酶。结合和未结合的 Fuc-α（1-6）-GlcNAc 的分子模拟，其中 GlcNAc 位于所有 N-聚糖的还原端，表明降低的 BKF 活性可能是由于在室温下，在 BKF 活性结合位点中最高占据构象的非最佳拟合。种群分析和自由能估计表明，标记糖的柔韧性增强，可通过延长反应条件来实现识别和结合。我们使用高浓度的 BKF 通过顺序外切糖苷酶消化阵列提供了这些实验条件。
