Rapid Labeling of Metabolically Engineered Cell-Surface Glycoconjugates with a Carbamate-Linked Cyclopropene Reporter

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• 作者：Anne-Katrin Sp?te ; Holger Bu?kamp ; Andrea Niederwieser ; Verena F. Schart ; Andreas Marx ; Valentin Wittmann
• 刊名：Bioconjugate Chemistry
• 出版年：2014
• 出版时间：January 15, 2014
• 年：2014
• 卷：25
• 期：1
• 页码：147-154
• 全文大小：579K
• ISSN：1520-4812

文摘

Metabolic oligosaccharide engineering is a valuable tool to monitor cellular carbohydrates. Here, we report the synthesis of a novel N-acyl-mannosamine derivative bearing a methylcyclopropene tag that is attached to the sugar via a carbamate moiety. This derivative undergoes rapid Diels鈥揂lder reaction with inverse electron demand. We demonstrate that the cell鈥檚 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鈥搕etrazine 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鈥揂lder reaction with inverse electron demand and a strain-promoted azide鈥揳lkyne cycloaddition which are carried out simultaneously in a single step.