TY - JOUR
T1 - Chemical Genetic Control of Protein Levels
T2 - Selective in Vivo Targeted Degradation
AU - Schneekloth, John S.
AU - Fonseca, Fabiana N.
AU - Koldobskiy, Michael
AU - Mandal, Amit
AU - Deshaies, Raymond
AU - Sakamoto, Kathleen
AU - Crews, Craig M.
PY - 2004/3/31
Y1 - 2004/3/31
N2 - Genetic loss of function analysis is a powerful method for the study of protein function. However, some cell biological questions are difficult to address using traditional genetic strategies often due to the lack of appropriate genetic model systems. Here, we present a general strategy for the design and syntheses of molecules capable of inducing the degradation of selected proteins in vivo via the ubiquitin-proteasome pathway. Western blot and fluorometric analyses indicated the loss of two different targets: green fluorescent protein (GFP) fused with FK506 binding protein (FKBP12) and GFP fused with the androgen receptor (AR), after treatment with PROteolysis TArgeting Chimeric molecules (PROTACS) incorporating a FKBP12 ligand and dihydrotestosterone, respectively. These are the first in vivo examples of direct small molecule-induced recruitment of target proteins to the proteasome for degradation upon addition to cultured cells. Moreover, PROTAC-mediated protein degradation offers a general strategy to create "chemical knockouts," thus opening new possibilities for the control of protein function.
AB - Genetic loss of function analysis is a powerful method for the study of protein function. However, some cell biological questions are difficult to address using traditional genetic strategies often due to the lack of appropriate genetic model systems. Here, we present a general strategy for the design and syntheses of molecules capable of inducing the degradation of selected proteins in vivo via the ubiquitin-proteasome pathway. Western blot and fluorometric analyses indicated the loss of two different targets: green fluorescent protein (GFP) fused with FK506 binding protein (FKBP12) and GFP fused with the androgen receptor (AR), after treatment with PROteolysis TArgeting Chimeric molecules (PROTACS) incorporating a FKBP12 ligand and dihydrotestosterone, respectively. These are the first in vivo examples of direct small molecule-induced recruitment of target proteins to the proteasome for degradation upon addition to cultured cells. Moreover, PROTAC-mediated protein degradation offers a general strategy to create "chemical knockouts," thus opening new possibilities for the control of protein function.
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U2 - 10.1021/ja039025z
DO - 10.1021/ja039025z
M3 - Article
C2 - 15038727
AN - SCOPUS:1642343326
SN - 0002-7863
VL - 126
SP - 3748
EP - 3754
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 12
ER -