Sialyltransferase Activity Assay for Ganglioside GM3 Synthase
Sialyltransferase Activity Assay for Ganglioside GM3 Synthase
复制标题
神经节苷脂 GM3 合酶的唾液酸转移酶活性测定
DOI：
10.1007/978-1-0716-2910-9_9
复制
发表时间：
2023
影响因子：
--
通讯作者：
Inokuchi Jin-ichi
中科院分区：
文献类型：
--
作者：
Inamori Kei-ichiro;Nitta Takahiro;Shishido Fumi;Watanabe Soichiro;Ohno Isao;Inokuchi Jin-ichi
GM 3合成酶（GM 3S）是一种唾液酸转移酶，将唾液酸从CMP-唾液酸转移到乳糖神经酰胺。该反应导致神经节苷脂GM 3的形成，并且对于其下游衍生物（包括a-和b-系列神经节苷脂）的生物合成是必需的。在这里，我们描述了一种使用荧光标记的烷基乳糖苷作为受体底物，然后通过HPLC分离酶产物的GM 3S酶测定方法。该方法允许定量测定培养细胞和小鼠脑组织中的GM 3S唾液酸转移酶活性。
GM3 synthase (GM3S) is a sialyltransferase that transfers sialic acid from CMP-sialic acid to lactosylceramide. This reaction results in formation of ganglioside GM3 and is essential for biosynthesis of its downstream derivatives, which include a- and b-series gangliosides. Here, we describe a method for GM3S enzymatic assay using fluorescence-labeled alkyl lactoside as acceptor substrate, followed by HPLC for separation of enzymatic product. The method allows quantitative assay of GM3S sialyltransferase activity in cultured cells and mouse brain tissues.