The ability to execute live-cell, localized glyco-editing is of pivotal importance for revolutionizing the field of glycobiology but no effective tool has been developed. We report herein the design of protein-specific glyco-editing probes, operating through localized enzyme decaging (LED), for live-cell editing of carbohydrate units on the protein of interest. The integration of aptamer recognition and molecular cloaking/uncloaking-based glyco-enzyme caging/decaging mechanism ensures the protein-specific editing. Proof-of-concept demonstration has been achieved for mucin 1 (MUC1)-specific terminal galactose/N-acetylgalactosamine (Gal/GalNAc) editing and sialic acid (Sia) trimming on live cells. A unique advantage of our LED strategy is the modular adaptability for performing cascade localized glyco-editing (CALOGE) manipulation through sequential aptamer docking-erasing-docking processes. Moreover, in vivo MUC1-specific Gal/GalNAc editing has also been realized, which enables the fluorescence visualization of Gal/GalNAc on MUC1 in murine bladder. In particular, specific trimming of MUC1-bound Sia on live cells has demonstrated, for the first time, the ability to directly reveal the significant effects of protein-specific glycoform on intracellular signaling and cell migration. These results suggest the probe as a generically applicable investigation tool for the revelation and manipulation of biological functions in association with protein-specific glycoforms.

执行活细胞、局部糖编辑的能力对于彻底改变糖生物学领域至关重要，但尚未开发出有效的工具。我们在此报告了蛋白质特异性糖编辑探针的设计，通过局部酶降解 (LED) 操作，用于对感兴趣蛋白质上的碳水化合物单位进行活细胞编辑。适配体识别和基于分子隐藏/解隐藏的糖酶笼罩/去壳机制的整合确保了蛋白质特异性编辑。已针对粘蛋白 1 (MUC1) 特异性末端半乳糖/ N实现了概念验证演示-乙酰半乳糖胺 (Gal/GalNAc) 编辑和唾液酸 (Sia) 修整活细胞。我们的 LED 策略的一个独特优势是通过顺序适体对接-擦除-对接过程执行级联局部糖编辑 (CALOGE) 操作的模块化适应性。此外，体内还实现了 MUC1 特定的 Gal/GalNAc 编辑，这使得 Gal/GalNAc 在小鼠膀胱中的 MUC1 上的荧光可视化成为可能。特别是，在活细胞上对结合 MUC1 的 Sia 进行特异性修剪，首次证明了直接揭示蛋白质特异性糖型对细胞内信号传导和细胞迁移的显着影响的能力。这些结果表明，该探针是一种普遍适用的调查工具，用于揭示和操纵与蛋白质特异性糖型相关的生物功能。