Metabolic delivery of ketone groups to sialic acid residues. Application to cell surface glycoform engineering

Kevin J Yarema, Lara K. Mahal, Richard E. Bruehl, Elena C. Rodriguez, Carolyn R. Bertozzi

Research output: Contribution to journalArticle

Abstract

The development of chemical strategies for decorating cells with defined carbohydrate epitopes would greatly facilitate studies of carbohydrate- mediated cell surface interactions. This report describes a general strategy for engineering the display of chemically defined oligosaccharides on cell surfaces that combines the concepts of metabolic engineering and selective chemical reactivity. Using a recently described method (Mahal, L. K., Yarema, K. J., and Bertozzi, C. R. (1997) Science 276, 1125-1128), we delivered a uniquely reactive ketone group to endogenous cell surface sialic acid residues by treating cells with the ketone-bearing metabolic precursor N- levulinoylmannosamine (ManLev). The ketone undergoes highly selective condensation reactions with complementary nucleophiles such as aminooxy and hydrazide groups. The detailed quantitative parameters of ManLev metabolism in human and nonhuman-derived cell lines were determined to establish a foundation for the modification of cell surfaces with novel epitopes at defined cell-surface densities. Ketones within the glycoconjugates on ManLev- treated cells were then reacted with synthetic aminooxy and hydrazide- functionalized carbohydrates. The remodeled cells were endowed with novel lectin binding profiles as determined by flow cytometry analysis. The simplicity and generality of this method make it well suited for use in the study of carbohydrate-mediated cell surface interactions.

Original languageEnglish (US)
Pages (from-to)31168-31179
Number of pages12
JournalJournal of Biological Chemistry
Volume273
Issue number47
DOIs
StatePublished - Nov 20 1998
Externally publishedYes

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N-Acetylneuraminic Acid
Ketones
Carbohydrates
Epitopes
Cell Communication
Bearings (structural)
Cells
Metabolic engineering
Chemical reactivity
Nucleophiles
Glycoconjugates
Condensation reactions
Flow cytometry
Metabolic Engineering
Oligosaccharides
Lectins
Metabolism
Display devices
Flow Cytometry
Cell Count

ASJC Scopus subject areas

  • Biochemistry

Cite this

Metabolic delivery of ketone groups to sialic acid residues. Application to cell surface glycoform engineering. / Yarema, Kevin J; Mahal, Lara K.; Bruehl, Richard E.; Rodriguez, Elena C.; Bertozzi, Carolyn R.

In: Journal of Biological Chemistry, Vol. 273, No. 47, 20.11.1998, p. 31168-31179.

Research output: Contribution to journalArticle

Yarema, Kevin J ; Mahal, Lara K. ; Bruehl, Richard E. ; Rodriguez, Elena C. ; Bertozzi, Carolyn R. / Metabolic delivery of ketone groups to sialic acid residues. Application to cell surface glycoform engineering. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 47. pp. 31168-31179.
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