N-Azidoacetylmannosamine-mediated chemical tagging of gangliosides
N-Azidoacetylmannosamine-mediated chemical tagging of gangliosides
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DOI：
10.1194/jlr.c700006-jlr200
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发表时间：
2007-06-01
影响因子：
6.5
通讯作者：
Overkleeft, Herman S.
中科院分区：
文献类型：
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作者：
Bussink, Anton P.;van Swieten, Paul F.;Overkleeft, Herman S.
过乙酰化n - α -叠氮酰基甘露胺（Ac(4)ManNAz）被细胞代谢为cmp -叠氮酰基甘露胺酸。先前已经证明，通过这种方式形成了含叠氮脲酸的糖蛋白，这些糖蛋白可以通过改良的Staudinger连接在细胞表面进行标记。在这里，我们首先证明了同样的过程也会导致含有叠氮脲酸的神经节苷脂的形成。脱氧麦诺吉里霉素是蛋白质中n -聚糖加工的抑制剂，可使Jurkat细胞的总细胞表面标记降低约25%。N-[5-（金刚烷-1-酰基甲氧基）-戊基]1-脱氧诺吉霉素抑制神经节苷脂生物合成可使细胞表面标记减少约75%。总之，细胞暴露于Ac(4)ManNAz可以在体内对神经节苷脂进行化学标记。
Peracetylated N-alpha-azidoacetylmannosamine (Ac(4)ManNAz) is metabolized by cells to CMP-azidosialic acid. It has been demonstrated previously that in this way azidosialic acid-containing glycoproteins are formed that can be labeled on the cell surface by a modified Staudinger ligation. Here, we first demonstrate that the same procedure also results in the formation of azidosialic acid-containing gangliosides. Deoxymannojirimycin, an inhibitor of N-glycan processing in proteins, decreases the total cell surface labeling in Jurkat cells by similar to 25%. Inhibition of ganglioside biosynthesis with N-[5-(adamantan-1-yl-methoxy)-pentyl]1-deoxynojirimycin reduces cell surface labeling by similar to 75%. In conclusion, exposure of cells to Ac(4)ManNAz allows in vivo chemical tagging of gangliosides.