3L95

Crystal structure of the human Notch1 Negative Regulatory Region (NRR) bound to the fab fragment of an antagonist antibody

Summary for 3L95

• Entry DOI: 10.2210/pdb3l95/pdb
• Related: 2OO4 3ETO
• Descriptor: anti-NRR1 fab fragment light chain, anti-NRR1 fab fragment heavy chain, Neurogenic locus notch homolog protein 1, ... (7 entities in total)
• Functional Keywords: nrr, fab fragment, antibody, alpha-beta-sandwich, sea domain, lnr, lin12 notch cysteine-rich, hd domain, cell cycle, signaling protein, activator, ank repeat, cell membrane, developmental protein, differentiation, egf-like domain, glycoprotein, membrane, metal-binding, notch signaling pathway, immune system
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 6
• Total formula weight: 150120.83

Authors

Hymowitz, S.G.,de Leon, G.P. (deposition date: 2010-01-04, release date: 2010-03-09, Last modification date: 2023-09-06)

Primary citation

Wu, Y.,Cain-Hom, C.,Choy, L.,Hagenbeek, T.J.,de Leon, G.P.,Chen, Y.,Finkle, D.,Venook, R.,Wu, X.,Ridgway, J.,Schahin-Reed, D.,Dow, G.J.,Shelton, A.,Stawicki, S.,Watts, R.J.,Zhang, J.,Choy, R.,Howard, P.,Kadyk, L.,Yan, M.,Zha, J.,Callahan, C.A.,Hymowitz, S.G.,Siebel, C.W.
Therapeutic antibody targeting of individual Notch receptors.
Nature, 464:1052-1057, 2010

PubMed Abstract: The four receptors of the Notch family are widely expressed transmembrane proteins that function as key conduits through which mammalian cells communicate to regulate cell fate and growth. Ligand binding triggers a conformational change in the receptor negative regulatory region (NRR) that enables ADAM protease cleavage at a juxtamembrane site that otherwise lies buried within the quiescent NRR. Subsequent intramembrane proteolysis catalysed by the gamma-secretase complex liberates the intracellular domain (ICD) to initiate the downstream Notch transcriptional program. Aberrant signalling through each receptor has been linked to numerous diseases, particularly cancer, making the Notch pathway a compelling target for new drugs. Although gamma-secretase inhibitors (GSIs) have progressed into the clinic, GSIs fail to distinguish individual Notch receptors, inhibit other signalling pathways and cause intestinal toxicity, attributed to dual inhibition of Notch1 and 2 (ref. 11). To elucidate the discrete functions of Notch1 and Notch2 and develop clinically relevant inhibitors that reduce intestinal toxicity, we used phage display technology to generate highly specialized antibodies that specifically antagonize each receptor paralogue and yet cross-react with the human and mouse sequences, enabling the discrimination of Notch1 versus Notch2 function in human patients and rodent models. Our co-crystal structure shows that the inhibitory mechanism relies on stabilizing NRR quiescence. Selective blocking of Notch1 inhibits tumour growth in pre-clinical models through two mechanisms: inhibition of cancer cell growth and deregulation of angiogenesis. Whereas inhibition of Notch1 plus Notch2 causes severe intestinal toxicity, inhibition of either receptor alone reduces or avoids this effect, demonstrating a clear advantage over pan-Notch inhibitors. Our studies emphasize the value of paralogue-specific antagonists in dissecting the contributions of distinct Notch receptors to differentiation and disease and reveal the therapeutic promise in targeting Notch1 and Notch2 independently.

PubMed: 20393564
DOI: 10.1038/nature08878

Experimental method

X-RAY DIFFRACTION (2.19 Å)