Abstract
Combined magnetic and fluorescence cell sorting were used to select Fluorogen Activating Proteins (FAPs) from a yeast surface-displayed library for binding to the fluorogenic cyanine dye Dimethyl Indole Red (DIR). Several FAPs were selected that bind to the dye with low nanomolar Kd values and enhance fluorescence more than 100-fold. One of these FAPs also exhibits considerable promiscuity, binding with high affinity to several other fluorogenic cyanine dyes with emission wavelengths covering most of the visible and near-IR regions of the spectrum. This significantly expands the number and wavelength range of scFv-based fluoromodules.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Carbocyanines / chemistry*
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Carbocyanines / metabolism
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Coloring Agents / chemistry
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Coloring Agents / metabolism
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Flow Cytometry / methods
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Fluorescent Dyes / chemistry*
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Fluorescent Dyes / metabolism
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Fungal Proteins / chemistry
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Immunoglobulin Fragments / chemistry*
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Immunoglobulin Fragments / metabolism
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Immunoglobulin Variable Region / chemistry*
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Immunoglobulin Variable Region / metabolism
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Microscopy, Fluorescence / methods
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Peptide Library
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Protein Binding
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Spectrometry, Fluorescence
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Spectrophotometry, Infrared
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Yeasts / chemistry
Substances
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Carbocyanines
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Coloring Agents
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Fluorescent Dyes
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Fungal Proteins
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Immunoglobulin Fragments
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Immunoglobulin Variable Region
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Peptide Library