以单分子灵敏度追踪活癌细胞上的表面聚糖。
Tracking surface glycans on live cancer cells with single-molecule sensitivity.
作者信息
Jiang Hao, English Brian P, Hazan Rachel B, Wu Peng, Ovryn Ben
机构信息
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461 (USA).
出版信息
Angew Chem Int Ed Engl. 2015 Feb 2;54(6):1765-9. doi: 10.1002/anie.201407976. Epub 2014 Dec 16.
Abstract
Using a combination of metabolically labeled glycans, a bioorthogonal copper(I)-catalyzed azide-alkyne cycloaddition, and the controlled bleaching of fluorescent probes conjugated to azide- or alkyne-tagged glycans, a sufficiently low spatial density of dye-labeled glycans was achieved, enabling dynamic single-molecule tracking and super-resolution imaging of N-linked sialic acids and O-linked N-acetyl galactosamine (GalNAc) on the membrane of live cells. Analysis of the trajectories of these dye-labeled glycans in mammary cancer cells revealed constrained diffusion of both N- and O-linked glycans, which was interpreted as reflecting the mobility of the glycan rather than to be caused by transient immobilization owing to spatial inhomogeneities on the plasma membrane. Stochastic optical reconstruction microscopy (STORM) imaging revealed the structure of dynamic membrane nanotubes.
摘要
通过结合代谢标记聚糖、生物正交铜（I）催化的叠氮化物-炔烃环加成反应以及对与叠氮化物或炔烃标记聚糖共轭的荧光探针进行可控漂白，实现了染料标记聚糖足够低的空间密度，从而能够对活细胞膜上的N-连接唾液酸和O-连接N-乙酰半乳糖胺（GalNAc）进行动态单分子追踪和超分辨率成像。对这些染料标记聚糖在乳腺癌细胞中的轨迹分析表明，N-连接和O-连接聚糖均存在受限扩散，这被解释为反映了聚糖的流动性，而非由质膜上的空间不均匀性导致的瞬时固定所致。随机光学重建显微镜（STORM）成像揭示了动态膜纳米管的结构。
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