Precise Control of Protein Concentration in Living Cells

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 (Pr...

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Published in:	Angewandte Chemie (International ed.) Vol. 49; no. 45; pp. 8458 - 8461
Main Authors:	Lau, Hubert D, Yaegashi, Junko, Zaro, Balyn W, Pratt, Matthew R
Format:	Journal Article
Language:	English
Published:	Weinheim Wiley‐VCH Verlag 02-11-2010 WILEY-VCH Verlag WILEY‐VCH Verlag
Subjects:	Amino Acid Substitution Animals Anti-Bacterial Agents - pharmacology destabilization domains Flow Cytometry Green Fluorescent Proteins - chemistry Green Fluorescent Proteins - metabolism Mice molecular recognition Mutagenesis, Site-Directed NIH 3T3 Cells protein concentration Protein Structure, Tertiary proteins Recombinant Fusion Proteins - chemistry Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Sirolimus - pharmacology split ubiquitin Tacrolimus Binding Proteins - chemistry Tacrolimus Binding Proteins - genetics Tacrolimus Binding Proteins - metabolism Ubiquitin - chemistry Ubiquitin - genetics Ubiquitin - metabolism
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Summary:	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.
Bibliography:	http://dx.doi.org/10.1002/anie.201003073 University of Southern California ark:/67375/WNG-MK1JB5ZH-T 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. istex:29B07B56142EACD943310B6171ED835A5D2909F7 ArticleID:ANIE201003073 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. ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1433-7851 1521-3773
DOI:	10.1002/anie.201003073