Get access

Precise Control of Protein Concentration in Living Cells

Authors

  • Hubert D. Lau,

    1. Departments of Chemistry and Molecular and Computational Biology, University of Southern California, 840 Downey Way, LJS 250, Los Angeles, CA 90089-0744 (USA), Fax: (+1) 213-740-0930
    Search for more papers by this author
  • Junko Yaegashi,

    1. Program in Biomedical and Biological Sciences, Keck School of Medicine, University of Southern California, 1975 Zonal Ave, KAM 110, Los Angeles, CA 90089-9023 (USA)
    Search for more papers by this author
  • Balyn W. Zaro,

    1. Departments of Chemistry and Molecular and Computational Biology, University of Southern California, 840 Downey Way, LJS 250, Los Angeles, CA 90089-0744 (USA), Fax: (+1) 213-740-0930
    Search for more papers by this author
  • Prof. Matthew R. Pratt

    1. Departments of Chemistry and Molecular and Computational Biology, University of Southern California, 840 Downey Way, LJS 250, Los Angeles, CA 90089-0744 (USA), Fax: (+1) 213-740-0930
    Search for more papers by this author

  • This research was supported by the University of Southern California. We also thank Dr. Lora Barsky in the USC-CSCRM/NCCC Flow Cytometry Core Facility, Prof. Debbie Johnson (USC) for tetracycline-responsive cells, Prof. Donald Arnold and Sarmad Al-Bassam (USC) for advice and instrumentation, Prof. Susan Lindquist (Whitehead) for α-synuclein cDNA, and Prof. Tom Muir (Rockefeller) for helpful discussions.

Abstract

original image

Rapamycin to the rescue: Native protein concentration was precisely controlled in living cells by using the small molecule rapamycin. The addition of rapamycin “shielded” a destabilization domain from degradation and induced complementation of split ubiquitin (UbN and UbC); as a result, proteins (Pro) of interest were released. This technology enables the investigation of native protein biology over a broad concentration range.

Ancillary