Department of Biochemistry, Albert Einstein College of Medicine, Yeshiva University, 1300 Morris Park Ave, Bronx, New York 10461, United States.
J Am Chem Soc. 2010 Dec 1;132(47):16893-9. doi: 10.1021/ja106553e. Epub 2010 Nov 9.
The Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) is the standard method for bioorthogonal conjugation. However, current Cu(I) catalyst formulations are toxic, hindering their use in living systems. Here we report that BTTES, a tris(triazolylmethyl)amine-based ligand for Cu(I), promotes the cycloaddition reaction rapidly in living systems without apparent toxicity. This catalyst allows, for the first time, noninvasive imaging of fucosylated glycans during zebrafish early embryogenesis. We microinjected embryos with alkyne-bearing GDP-fucose at the one-cell stage and detected the metabolically incorporated unnatural sugars using the biocompatible click chemistry. Labeled glycans could be imaged in the enveloping layer of zebrafish embryos between blastula and early larval stages. This new method paves the way for rapid, noninvasive imaging of biomolecules in living organisms.
铜（I）催化的叠氮化物-炔烃环加成（CuAAC）是生物正交共轭的标准方法。然而，目前的铜（I）催化剂配方具有毒性，阻碍了它们在活系统中的应用。在这里，我们报告说，BTTES，一种基于三（三唑基甲基）胺的铜（I）配体，在活系统中快速促进环加成反应，而没有明显的毒性。这种催化剂首次允许在斑马鱼早期胚胎发生过程中非侵入性地成像岩藻糖基化聚糖。我们在单细胞阶段将带有炔基的 GDP-岩藻糖微注射到胚胎中，并使用生物相容性的点击化学检测代谢掺入的非天然糖。在胚胎的胚泡和早期幼虫阶段之间的包被层中可以对标记的聚糖进行成像。这种新方法为在活生物体中快速、非侵入性地成像生物分子铺平了道路。
