5CZV

Crystal structure of Notch3 NRR in complex with 20350 Fab

Summary for 5CZV

• Entry DOI: 10.2210/pdb5czv/pdb
• Related: 5CZX
• Descriptor: Neurogenic locus notch homolog protein 3, Fab 20350 heavy chain, Fab 20350 light chain, ... (6 entities in total)
• Functional Keywords: antibody, notch3, oncology, immune system
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 3
• Total formula weight: 77789.93

Authors

Hu, T.,Fryer, C.,Chopra, R.,Clark, K. (deposition date: 2015-08-01, release date: 2016-06-01, Last modification date: 2024-10-16)

Primary citation

Bernasconi-Elias, P.,Hu, T.,Jenkins, D.,Firestone, B.,Gans, S.,Kurth, E.,Capodieci, P.,Deplazes-Lauber, J.,Petropoulos, K.,Thiel, P.,Ponsel, D.,Hee Choi, S.,LeMotte, P.,London, A.,Goetcshkes, M.,Nolin, E.,Jones, M.D.,Slocum, K.,Kluk, M.J.,Weinstock, D.M.,Christodoulou, A.,Weinberg, O.,Jaehrling, J.,Ettenberg, S.A.,Buckler, A.,Blacklow, S.C.,Aster, J.C.,Fryer, C.J.
Characterization of activating mutations of NOTCH3 in T-cell acute lymphoblastic leukemia and anti-leukemic activity of NOTCH3 inhibitory antibodies.
Oncogene, 35:6077-6086, 2016

PubMed Abstract: Notch receptors have been implicated as oncogenic drivers in several cancers, the most notable example being NOTCH1 in T-cell acute lymphoblastic leukemia (T-ALL). To characterize the role of activated NOTCH3 in cancer, we generated an antibody that detects the neo-epitope created upon gamma-secretase cleavage of NOTCH3 to release its intracellular domain (ICD3), and sequenced the negative regulatory region (NRR) and PEST (proline, glutamate, serine, threonine) domain coding regions of NOTCH3 in a panel of cell lines. We also characterize NOTCH3 tumor-associated mutations that result in activation of signaling and report new inhibitory antibodies. We determined the structural basis for receptor inhibition by obtaining the first co-crystal structure of a NOTCH3 antibody with the NRR protein and defined two distinct epitopes for NRR antibodies. The antibodies exhibit potent anti-leukemic activity in cell lines and tumor xenografts harboring NOTCH3 activating mutations. Screening of primary T-ALL samples reveals that 2 of 40 tumors examined show active NOTCH3 signaling. We also identified evidence of NOTCH3 activation in 12 of 24 patient-derived orthotopic xenograft models, 2 of which exhibit activation of NOTCH3 without activation of NOTCH1. Our studies provide additional insights into NOTCH3 activation and offer a path forward for identification of cancers that are likely to respond to therapy with NOTCH3 selective inhibitory antibodies.

PubMed: 27157619
DOI: 10.1038/onc.2016.133

Experimental method

X-RAY DIFFRACTION (3.19 Å)