Enhanced photostability of genetically encodable fluoromodules based on fluorogenic cyanine dyes and a promiscuous protein partner

J Am Chem Soc. 2009 Sep 16;131(36):12960-9. doi: 10.1021/ja9016864.

Abstract

Fluoromodules are discrete complexes of biomolecules and fluorogenic dyes. Binding of the dyes to their cognate biomolecule partners results in enhanced dye fluorescence. We exploited a previously reported promiscuous binding interaction between a single-chain, variable fragment antibody protein and a family of cyanine dyes to create new protein-dye fluoromodules that exhibit enhanced photostability while retaining high affinity protein-dye binding. Modifications to the dye structure included electron-withdrawing groups that provide resistance to photo-oxidative damage. Low nanomolar equilibrium dissociation constants were found for the new dyes. Fluorescence microscopy illustrates how yeast can be surface-labeled with three different colors based on a single protein and appropriately chosen dyes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites, Antibody
  • Carbocyanines / analysis
  • Carbocyanines / metabolism*
  • Fluorescent Dyes / analysis
  • Fluorescent Dyes / metabolism*
  • Immunoglobulin Fab Fragments / genetics
  • Immunoglobulin Fab Fragments / metabolism*
  • Microscopy, Fluorescence
  • Molecular Structure
  • Photochemistry
  • Protein Binding
  • Saccharomyces cerevisiae / cytology

Substances

  • Carbocyanines
  • Fluorescent Dyes
  • Immunoglobulin Fab Fragments