1 research outputs found
Rapid Labeling of Metabolically Engineered Cell-Surface Glycoconjugates with a Carbamate-Linked Cyclopropene Reporter
Metabolic oligosaccharide engineering
is a valuable tool to monitor
cellular carbohydrates. Here, we report the synthesis of a novel <i>N</i>-acyl-mannosamine derivative bearing a methylcyclopropene
tag that is attached to the sugar via a carbamate moiety. This derivative
undergoes rapid Diels–Alder reaction with inverse electron
demand. We demonstrate that the cell’s biosynthetic machinery
incorporates this non-natural mannosamine derivative into glycoconjugates
that can, subsequently, be labeled within less than 10 min with a
new sulfo-Cy3–tetrazine conjugate. Using this tetrazine-dye
conjugate for the detection of the methylcyclopropene-tagged mannosamine
derivative, we could achieve dual labeling of two different metabolically
incorporated sugars combining a Diels–Alder reaction with inverse
electron demand and a strain-promoted azide–alkyne cycloaddition
which are carried out simultaneously in a single step