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Probing Protein–Protein Interactions with a Genetically Encoded Photo-crosslinking Amino Acid

Authors

  • Dr. Hui-wang Ai,

    1. Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
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    • These authors contributed equally to this work.

  • Dr. Weijun Shen,

    1. Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
    2. Present address: Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA 92121 (USA)
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    • These authors contributed equally to this work.

  • Dr. Amit Sagi,

    1. Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
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  • Dr. Peng R. Chen,

    1. Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
    2. Present address: Department of Chemical Biology, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871 (China)
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  • Prof. Dr. Peter G. Schultz

    Corresponding author
    1. Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
    • Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
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Abstract

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A pyrrolysyl tRNA/tRNA sythetase pair has been evolved that efficiently incorporates an aliphatic diazirine amino acid into proteins in E. coli and mammalian cells. When site-specifically introduced into the active site of Cdk5, this unnatural amino acid was able to photo-crosslink the kinase to its substrate, Pak1.

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