Cu-free Click Chemistry In Glycan Imaging
hhmi John C. Jewett, Ellen M. Sletten, and Carolyn R. Bertozzi
University of California, Berkeley, California 94720
Abstract
Bioorthogonal reactions are chemical reactions that neither interact
with nor interfere with a biological system. The participating
functional groups must be inert to biological moieties, must
selectively react with each other under biocompatible conditions,
Carolyn Bertozzi modified the original click reaction to produce a
copper-free version. She used this reaction to track molecules called
glycans on cell surfaces, which she had been investigating since the
early 1990s.
Background
when cells undergo a transformation of their state there are changes
in the patterns of cell surface glycans and sometimes those changes in
cell surface glycosylation are associated with disease for example it
was already known back then that when healthy cells transform into a
cancerous state in the context of a tumor there are cell surface
glycosylation changes that often include an overproduction of a
particular sugar that's known as sialic acid and it's one of the building
blocks in the complex carbohydrates on the surface of cells
Fakher NTICHA
Materials and Methods Results
We envisioned a two-step process for glycan imaging: We sought to evaluate BARAC’s performance in live cell
(1) Metabolic labeling, and (2) bioorthogonal chemistry
labeling experiments. Using a carbamate linkage.
 Even at a concentration of 50 nM, BARAC-biotin still
showed significant cell labeling in 1 h
 The high level of reactivity of BARAC-biotin was not
accompanied by any cytotoxicity compared to cells
treated with no cyclooctyne reagent
X = Azide
Y = Cyclooctyne
In designing new cyclooctyne
probes we sought to brush against the line between
stability and reactivity without crossing it.
Conclusion
BARAC is therefore a promising reagent for real-time and in
vivo imaging of azide-labeled biomolecules , thus a great tool
to explore more of the vertebrate biology.
There are no reported cyclooctynes that have an amide Future Direction
within the ring, as in BARAC; thus, their balance of stability
and reactivity is an intriguing unanswered question. We
Nowadays our straightforward goal is to develop
studied the reactivity of BARAC using benzyl azide as a
bioorthogonal reactions with even higher selectivity and
model substrate. Gratifyingly, the second-order rate constant
orthogonality that would allow for tracking glycans clinically.
in acetonitrile at rt was 0.96 𝑀 𝑠 over 12-fold higher
than the rate constant for DIFO under identical conditions
Acknowledgments
This study was supported by a postdoctoral fellowship from the American Cancer
Society, and E.M.S. was supported by a predoctoral fellowship from the Organic
Division of the American Chemical Society (Genentech). This work was
supported by a grant to C.R.B. from the National Institutes of Health (GM58867).