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Novel assay for direct fluorescent imaging of sialidase activity

机译：用于唾液酸酶活性直接荧光成像的新型测定

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Here we describe a novel approach to sialidase activity estimation. Sialidases (EC 3.2.1.18, exo-α-sialidases), also known as neuraminidases, are the group of enzymes, which hydrolyze the glycoside bound between terminal sialic acid and subsequent carbohydrate residue in glycoproteins and glycolipids. Sialic acids are the group of monosaccharides with acidic properties, since they are acetylated or glycolylated derivates of neuraminic acid. Flu and some other viruses use neuraminidase activity to infect host cells. The level of sialylation was shown to be tightly connected with tumor cell invasiveness and metastatic potential, sialylation level also determines the clearance of aged or virus-infected cells. Thus, detection of sialidase activity is of primary importance for clinical diagnostics as well as life science research. The authors developed the assay for both visualization and estimation of sialidase activity in living cells. Previously known methods for sialidase activity detection required destruction of cellular material, or were low-sensitive, or provided no information on the activity localization in certain intracellular compartment. To overcome these problems, a fluorogenic neuraminidase substrate, 4-MUNA was utilized, and the method for detection of neuraminidase activity using fluorescent microscopy was proposed, it provided a high signal level and information on cellular localization of the studied enzyme. By using this approach the increase of sialidase activity on apoptotic cells was demonstrated in comparison to viable and primary necrotic cells
机译：在这里，我们描述了一种新的唾液酸酶活性估计方法。 Sialidase（EC 3.2.1.18，EXO-α-唾液酸酶）也称为神经氨酸酶，是酶组，其水解糖蛋白和糖脂蛋白在糖蛋白和糖脂的后续碳水化合物残留物之间的糖苷。唾液酸是具有酸性特性​​的单糖组，因为它们是乙酰化的或糖化衍生物的神经酸的衍生物。流感和其他一些病毒使用神经氨酸酶活性来感染宿主细胞。显示唾液酸化水平与肿瘤细胞侵袭性和转移潜力紧密相关，唾液酸化水平也决定了老年或病毒感染细胞的间隙。因此，检测唾液酸酶活性对于临床诊断以及生命科学研究的主要重要性。作者开发了活细胞中唾液酸酶活性的可视化和估算的测定。以前已知的唾液酸酶活性检测方法需要破坏细胞材料，或者是低敏感的，或者不提供关于某些细胞内隔室中的活性定位的信息。为了克服这些问题，提出了一种荧光神经氨酸酶底物，4-MUNA，以及使用荧光显微镜检测神经氨酸酶活性的方法，提供了高信号水平和关于研究酶细胞定位的信息。通过使用这种方法，与可行和原代坏死细胞相比，对凋亡细胞的唾液酸酶活性的增加

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