TY - JOUR
T1 - Modulation of notch signaling by antibodies specific for the extracellular negative regulatory region of NOTCH3
AU - Li, Kang
AU - Li, Yucheng
AU - Wu, Wenjuan
AU - Gordon, Wendy R.
AU - Chang, David W.
AU - Lu, Mason
AU - Scoggin, Shane
AU - Fu, Tihui
AU - Vien, Long
AU - Histen, Gavin
AU - Zheng, Ji
AU - Martin-Hollister, Rachel
AU - Duensing, Thomas
AU - Singh, Sanjaya
AU - Blacklow, Stephen C.
AU - Yao, Zhengbin
AU - Aster, Jon C.
AU - Zhou, Bin Bing S.
PY - 2008/3/21
Y1 - 2008/3/21
N2 - The Notch pathway regulates the development of many tissues and cell types and is involved in a variety of human diseases, making it an attractive potential therapeutic target. This promise has been limited by the absence of potent inhibitors or agonists that are specific for individual human Notch receptors (NOTCH1-4). Using an unbiased functional screening, we identified monoclonal antibodies that specifically inhibit or induce activating proteolytic cleavages in NOTCH3. Remarkably, the most potent inhibitory and activating antibodies bind to overlapping epitopes within a juxtamembrane negative regulatory region that protects NOTCH3 from proteolysis and activation in its resting auto-inhibited state. The inhibitory antibodies revert phenotypes conveyed on 293T cells by NOTCH3 signaling, such as increased cellular proliferation, survival, and motility, whereas the activating antibody mimics some of the effects of ligand-induced Notch activation. These findings provide insights into the mechanisms of Notch autoinhibition and activation and pave the way for the further development of specific antibody-based modulators of the Notch receptors, which are likely to be of utility in a wide range of experimental and therapeutic settings.
AB - The Notch pathway regulates the development of many tissues and cell types and is involved in a variety of human diseases, making it an attractive potential therapeutic target. This promise has been limited by the absence of potent inhibitors or agonists that are specific for individual human Notch receptors (NOTCH1-4). Using an unbiased functional screening, we identified monoclonal antibodies that specifically inhibit or induce activating proteolytic cleavages in NOTCH3. Remarkably, the most potent inhibitory and activating antibodies bind to overlapping epitopes within a juxtamembrane negative regulatory region that protects NOTCH3 from proteolysis and activation in its resting auto-inhibited state. The inhibitory antibodies revert phenotypes conveyed on 293T cells by NOTCH3 signaling, such as increased cellular proliferation, survival, and motility, whereas the activating antibody mimics some of the effects of ligand-induced Notch activation. These findings provide insights into the mechanisms of Notch autoinhibition and activation and pave the way for the further development of specific antibody-based modulators of the Notch receptors, which are likely to be of utility in a wide range of experimental and therapeutic settings.
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U2 - 10.1074/jbc.M800170200
DO - 10.1074/jbc.M800170200
M3 - Article
C2 - 18182388
AN - SCOPUS:43149113910
SN - 0021-9258
VL - 283
SP - 8046
EP - 8054
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 12
ER -