Cell surface engineering by a modified Staudinger reaction

Science. 2000 Mar 17;287(5460):2007-10. doi: 10.1126/science.287.5460.2007.

Abstract

Selective chemical reactions enacted within a cellular environment can be powerful tools for elucidating biological processes or engineering novel interactions. A chemical transformation that permits the selective formation of covalent adducts among richly functionalized biopolymers within a cellular context is presented. A ligation modeled after the Staudinger reaction forms an amide bond by coupling of an azide and a specifically engineered triarylphosphine. Both reactive partners are abiotic and chemically orthogonal to native cellular components. Azides installed within cell surface glycoconjugates by metabolism of a synthetic azidosugar were reacted with a biotinylated triarylphosphine to produce stable cell-surface adducts. The tremendous selectivity of the transformation should permit its execution within a cell's interior, offering new possibilities for probing intracellular interactions.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acetylation
  • Azides / chemical synthesis
  • Azides / chemistry
  • Azides / metabolism*
  • Biotin / analogs & derivatives*
  • Biotin / chemical synthesis
  • Biotin / chemistry
  • Biotin / metabolism
  • Biotinylation
  • Cell Membrane / chemistry*
  • Cell Membrane / metabolism*
  • Flow Cytometry
  • Fluorescent Dyes / metabolism
  • Glycoconjugates / metabolism
  • HeLa Cells
  • Hexosamines / chemical synthesis
  • Hexosamines / chemistry
  • Hexosamines / metabolism*
  • Humans
  • Jurkat Cells
  • Ketones / metabolism
  • Oxidation-Reduction
  • Phosphines / chemical synthesis
  • Phosphines / chemistry
  • Phosphines / metabolism*
  • Solubility
  • Sulfhydryl Compounds / metabolism

Substances

  • 4-(2-(2-(2-(biotinylamino)ethoxy)ethoxy)ethoxy)aminocarbonyl-2-diphenylphosphinobenzoic acid
  • Azides
  • Fluorescent Dyes
  • Glycoconjugates
  • Hexosamines
  • Ketones
  • N-azidoacetylmannosamine
  • Phosphines
  • Sulfhydryl Compounds
  • Biotin