Enhanced Photostability of Genetically Encodable Fluoromodules Based on Fluorogenic Cyanine Dyes and a Promiscuous Protein Partner

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• 作者：Nathaniel I. Shank ; Kimberly J. Zanotti ; Frederick Lanni ; Peter B. Berget ; Bruce A. Armitage
• 刊名：Journal of the American Chemical Society
• 出版年：2009
• 出版时间：September 16, 2009
• 年：2009
• 卷：131
• 期：36
• 页码：12960-12969
• 全文大小：461K
• 年卷期：v.131,no.36(September 16, 2009)
• ISSN：1520-5126

文摘

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.