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Proximity Enzymatic Glyco-Remodeling Enables Direct and Highly Efficient Lipid Raft Imaging on Live Cells
journal contribution
posted on 2020-04-28, 21:03 authored by Jing Tao, Xiaofei Yu, Yuna Guo, Guyu Wang, Huangxian Ju, Lin DingLipid
rafts, highly ordered cell membrane domains mainly composed
of cholesterol, sphingolipids, and protein receptors, serve as important
functional platforms for regulation of lipid/protein interactions.
The major predicament in lipid raft study is the lack of direct and
robust visualization tools for in situ tracking raft components. To
solve this issue, we herein report a proximity enzymatic glyco-remodeling
strategy for direct and highly efficient lipid raft labeling and imaging
on live cells. Through cofunctionalization of raft-specific recognition
motif and glycan-remodeling enzyme on gold nanoparticles, the fabricated
nanoprobe can be specifically guided to the raft domains to perform
catalytic remodeling on neighboring glycans. Taking advantage of the
abundant glycoconjugates enriched in lipid rafts, this elaborate design
achieves the translation of one raft-recognition event to multiple
raft-confined labeling operations, thus, significantly increasing
the labeling efficiency and imaging sensitivity. The direct covalent
labeling also enables in situ and long-term tracking of raft components
in live cells. The method possesses broad applicability and potential
expansibility, thus, will greatly facilitate the investigations on
the complex composition, organization, and dynamics of lipid rafts.