Metabolic labeling of fucosylated glycans in developing zebrafish.
Metabolic labeling of fucosylated glycans in developing zebrafish.
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DOI：
10.1021/cb100284d
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发表时间：
2011-06-17
影响因子：
4
通讯作者：
Bertozzi, Carolyn R.
中科院分区：
文献类型：
--
作者：
Dehnert, Karen W.;Beahm, Brendan J.;Huynh, Thinh T.;Baskin, Jeremy M.;Laughlin, Scott T.;Wang, Wei;Wu, Peng;Amacher, Sharon L.;Bertozzi, Carolyn R.
许多发育过程依赖于适当的岩藻糖基化，但这种翻译后修饰在体内难以监测。在此，我们应用一种化学报告分子策略来观察发育中的斑马鱼体内的岩藻糖基化聚糖。利用叠氮化物衍生的岩藻糖类似物，我们对细胞表面聚糖进行代谢标记，然后通过与二氟代环辛炔探针的无铜点击化学来检测掺入的叠氮化物。我们发现岩藻糖补救途径的酶在斑马鱼胚胎发生过程中表达，但它们对叠氮化物修饰的底物处理效率低下。我们能够通过使用一种叠氮化物官能化的GDP - 岩藻糖类似物来绕过补救途径。这种核苷糖很容易被岩藻糖基转移酶接受，并为岩藻糖基化聚糖提供了强有力的细胞表面标记，这通过流式细胞术和共聚焦显微镜分析得以确定。我们利用这项技术对斑马鱼胚胎发育前5天的包被层中的岩藻糖基化聚糖进行成像。这项工作提供了一种在体内研究岩藻糖基化聚糖生物合成的方法。
Many developmental processes depend on proper fucosylation, but this post-translational modification is difficult to monitor in vivo. Here we applied a chemical reporter strategy to visualize fucosylated glycans in developing zebrafish. Using azide-derivatized analogues of fucose, we metabolically labeled cell-surface glycans and then detected the incorporated azides via copper-free click chemistry with a difluorinated cyclooctyne probe. We found that the fucose salvage pathway enzymes are expressed during zebrafish embryogenesis but that they process the azide-modified substrates inefficiently. We were able to bypass the salvage pathway by using an azide-functionalized analogue of GDP-fucose. This nucleotide sugar was readily accepted by fucosyltransferases and provided robust cell-surface labeling of fucosylated glycans, as determined by flow cytometry and confocal microscopy analysis. We used this technique to image fucosylated glycans in the enveloping layer of zebrafish embryos during the first 5 days of development. This work provides a method to study the biosynthesis of fucosylated glycans in vivo.
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影响因子：
46.2
作者：
Jewett JC;Bertozzi CR
通讯作者：
Bertozzi CR
影响因子：
4.8
作者：
Roos, C;Kolmer, M;Renkonen, R
通讯作者：
Renkonen, R
影响因子：
15
作者：
Rabuka, David;Hubbard, Sarah C.;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.
DOI：
10.1073/pnas.0605418103
发表时间：
2006-08-15
影响因子：
11.1
作者：
Sawa, Masaaki;Hsu, Tsui-Ling;Wong, Chi-Huey
通讯作者：
Wong, Chi-Huey
DOI：
10.1083/jcb.200203125
发表时间：
2002-08-19
期刊：
The Journal of cell biology
影响因子：
--
作者：
Smith PL;Myers JT;Rogers CE;Zhou L;Petryniak B;Becker DJ;Homeister JW;Lowe JB
通讯作者：
Lowe JB