Ganglioside glycosyltransferase assay using ion-exchange chromatography.
Ganglioside glycosyltransferase assay using ion-exchange chromatography.
复制标题
使用离子交换色谱法进行神经节苷脂糖基转移酶测定。
DOI：
10.1016/0003-2697(86)90134-x
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发表时间：
1986
影响因子：
2.9
通讯作者：
Schnaar,RL
中科院分区：
文献类型：
--
作者：
Walton,KM;Schnaar,RL
采用离子交换色谱法开发了一种快速测定神经节苷脂糖基转移酶活性的方法。使用 13 天的鸡脑作为尿苷二磷酸-N-乙酰基-d-半乳糖胺：神经节苷脂 GM3N-乙酰半乳糖胺基转移酶（神经节苷脂 GM2 合成酶）的来源，我们能够通过将反应混合物应用到小柱中，准确测量 N-[3H]乙酰半乳糖胺 (GalNAc) 从 UDP-[3H]GalNAc 到 GM3 的转移。 DEAE-Sepharose 并用 10 mm 乙酸钾的甲醇溶液洗脱放射性标记的 GM2 反应产物。事实证明，该方法与之前发表的检测方法一样可靠且灵敏，但需要的时间和操作更少。
A rapid method for determination of ganglioside glycosyltransferase activity has been developed employing ion-exchange chromatography. Using 13-day chick brain as a source of uridine diphospho-N-acetyl-d-galactosamine: ganglioside GM3N-acetylgalactosaminyltransferase (ganglioside GM2synthetase), we were able to accurately measure transfer of N-[3H]acetylgalactosamine (GalNAc) from UDP-[3H]GalNAc to GM3by application of the reaction mixture to small columns of DEAE-Sepharose and elution of radiolabeled GM2reaction product with 10 mm potassium acetate in methanol. This method proved to be as reliable and sensitive as previously published assays but requires less time and fewer manipulations.