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

Nathaniel I. Shank, Kimberly J. Zanotti, Frederick Lanni, Peter B. Berget, Bruce A. Armitage

Research output: Contribution to journalArticlepeer-review

78 Scopus citations

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 electronwithdrawing 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.

Original languageEnglish
Pages (from-to)12960-12969
Number of pages10
JournalJournal of the American Chemical Society
Volume131
Issue number36
DOIs
StatePublished - Sep 16 2009

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