Copper-free click chemistry for dynamic in vivo imaging

Dynamic imaging of proteins in live cells is routinely performed by using genetically encoded reporters, an approach that cannot be extended to other classes of biomolecules such as glycans and lipids. Here, we report a Cu-free variant of click chemistry that can label these biomolecules rapidly and...

Full description

Saved in:
Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2007-10, Vol.104 (43), p.16793-16797
Main Authors: Baskin, Jeremy M, Prescher, Jennifer A, Laughlin, Scott T, Agard, Nicholas J, Chang, Pamela V, Miller, Isaac A, Lo, Anderson, Codelli, Julian A, Bertozzi, Carolyn R
Format: Article
Language:English
Subjects:
Animals
Azide
Azides
Biochemistry
Biological Sciences
Biological Transport
Biomolecules
Carbohydrates
Catalysis
Cell Survival
Chemical reactions
Chemistry, Organic - methods
CHO Cells
Click chemistry
Copper
Copper - metabolism
Cricetinae
Cricetulus
Cycloaddition
Endocytosis
Humans
Hydrocarbons, Cyclic - chemical synthesis
Hydrocarbons, Cyclic - chemistry
Hydrocarbons, Fluorinated - chemical synthesis
Hydrocarbons, Fluorinated - chemistry
Imaging
Imaging, Three-Dimensional - methods
Jurkat Cells
Kinetics
Ligation
Lipids
Molecular structure
Physical Sciences
Polysaccharides
Polysaccharides - metabolism
Proteins
Proteins - metabolism
Reagents
Scientific imaging
T lymphocytes
Time Factors
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Dynamic imaging of proteins in live cells is routinely performed by using genetically encoded reporters, an approach that cannot be extended to other classes of biomolecules such as glycans and lipids. Here, we report a Cu-free variant of click chemistry that can label these biomolecules rapidly and selectively in living systems, overcoming the intrinsic toxicity of the canonical Cu-catalyzed reaction. The critical reagent, a substituted cyclooctyne, possesses ring strain and electron-withdrawing fluorine substituents that together promote the [3 + 2] dipolar cycloaddition with azides installed metabolically into biomolecules. This Cu-free click reaction possesses comparable kinetics to the Cu-catalyzed reaction and proceeds within minutes on live cells with no apparent toxicity. With this technique, we studied the dynamics of glycan trafficking and identified a population of sialoglycoconjugates with unexpectedly rapid internalization kinetics.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0707090104