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A General Chemical Method to Regulate Protein Stability in the Mammalian Central Nervous System

Iwamoto, Mari; Björklund, Tomas LU ; Lundberg, Cecilia LU ; Kirik, Deniz LU and Wandless, Thomas J. (2010) In Chemistry and Biology 17(9). p.981-988
Abstract
The ability to make specific perturbations to biological molecules in a cell or organism is a central experimental strategy in modern research biology. We have developed a general technique in which the stability of a specific protein is regulated by a cell-permeable small molecule. Mutants of the Escherichia coil dihydrofolate reductase (ecDHFR) were engineered to be degraded, and, when this destabilizing domain is fused to a protein of interest, its instability is conferred to the fused protein resulting in rapid degradation of the entire fusion protein. A small-molecule ligand trimethoprim (TMP) stabilizes the destabilizing domain in a rapid, reversible, and dose-dependent manner, and protein levels in the absence of TMP are barely... (More)
The ability to make specific perturbations to biological molecules in a cell or organism is a central experimental strategy in modern research biology. We have developed a general technique in which the stability of a specific protein is regulated by a cell-permeable small molecule. Mutants of the Escherichia coil dihydrofolate reductase (ecDHFR) were engineered to be degraded, and, when this destabilizing domain is fused to a protein of interest, its instability is conferred to the fused protein resulting in rapid degradation of the entire fusion protein. A small-molecule ligand trimethoprim (TMP) stabilizes the destabilizing domain in a rapid, reversible, and dose-dependent manner, and protein levels in the absence of TMP are barely detectable. The ability of TMP to cross the blood-brain barrier enables the tunable regulation of proteins expressed in the mammalian central nervous system. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Chemistry and Biology
volume
17
issue
9
pages
981 - 988
publisher
Cell Press
external identifiers
  • wos:000283283200012
  • scopus:77956972260
ISSN
1879-1301
DOI
10.1016/j.chembiol.2010.07.009
language
English
LU publication?
yes
id
f2612416-f77d-4725-9fac-4e73c9473e7c (old id 1721006)
date added to LUP
2010-12-15 14:01:07
date last changed
2018-07-01 03:00:56
@article{f2612416-f77d-4725-9fac-4e73c9473e7c,
  abstract     = {The ability to make specific perturbations to biological molecules in a cell or organism is a central experimental strategy in modern research biology. We have developed a general technique in which the stability of a specific protein is regulated by a cell-permeable small molecule. Mutants of the Escherichia coil dihydrofolate reductase (ecDHFR) were engineered to be degraded, and, when this destabilizing domain is fused to a protein of interest, its instability is conferred to the fused protein resulting in rapid degradation of the entire fusion protein. A small-molecule ligand trimethoprim (TMP) stabilizes the destabilizing domain in a rapid, reversible, and dose-dependent manner, and protein levels in the absence of TMP are barely detectable. The ability of TMP to cross the blood-brain barrier enables the tunable regulation of proteins expressed in the mammalian central nervous system.},
  author       = {Iwamoto, Mari and Björklund, Tomas and Lundberg, Cecilia and Kirik, Deniz and Wandless, Thomas J.},
  issn         = {1879-1301},
  language     = {eng},
  number       = {9},
  pages        = {981--988},
  publisher    = {Cell Press},
  series       = {Chemistry and Biology},
  title        = {A General Chemical Method to Regulate Protein Stability in the Mammalian Central Nervous System},
  url          = {http://dx.doi.org/10.1016/j.chembiol.2010.07.009},
  volume       = {17},
  year         = {2010},
}