Unnatural Amino Acid Mutagenesis of Fluorescent Proteins

Authors

  • Dr. Feng Wang,

    1. Department of Chemistry, The Skaggs Institute for Chemical Biology, 10550 North Torrey Pines Road, La Jolla, CA 92037 (USA)
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  • Dr. Wei Niu,

    1. Department of Chemistry, University of Nebraska – Lincoln, Lincoln, NE 68588 (USA)
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  • Prof. Jiantao Guo,

    Corresponding author
    1. Department of Chemistry, University of Nebraska – Lincoln, Lincoln, NE 68588 (USA)
    • Department of Chemistry, University of Nebraska – Lincoln, Lincoln, NE 68588 (USA)
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  • Prof. Peter G. Schultz

    Corresponding author
    1. Department of Chemistry, The Skaggs Institute for Chemical Biology, 10550 North Torrey Pines Road, La Jolla, CA 92037 (USA)
    • Department of Chemistry, The Skaggs Institute for Chemical Biology, 10550 North Torrey Pines Road, La Jolla, CA 92037 (USA)
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  • This work is supported by grant DE-FG03-00ER46051 from The Division of Materials Sciences, DOE (P.G.S.), the Skaggs Institute for Chemical Biology (P.G.S.), and the New Faculty Startup Fund to J.G. from the Chemistry Department of University of Nebraska – Lincoln.

Abstract

original image

Tyrosine 66 of a green fluorescent protein (GFP) was substituted with unnatural amino acids carrying boronate, azido, nitro, and keto substituents. In general, the equation imagevalues of these GFP mutants is blue-shifted relative to that of GFP, and the fluorescence intensity of the boronate variant increases upon oxidation (see scheme). The X-ray crystal structures of the keto and boronate GFP mutants provide explanations of their altered fluorescence properties.

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