3ETO

2 Angstrom Xray structure of the NOTCH1 Negative Regulatory Region (NRR)

Summary for 3ETO

• Entry DOI: 10.2210/pdb3eto/pdb
• Descriptor: Neurogenic locus notch homolog protein 1, CALCIUM ION, CHLORIDE ION, ... (5 entities in total)
• Functional Keywords: alpha-beta sandwich, hd domain, lnr repeat, calcium-binding, sea domain, autoinhibition, activator, t-all, leukemia, oncogene, ank repeat, developmental protein, differentiation, egf-like domain, glycoprotein, membrane, metal-binding, notch signaling pathway, nucleus, phosphoprotein, receptor, transcription, transcription regulation, transmembrane, signaling protein
• Biological source: Homo sapiens
• Cellular location: Cell membrane; Single-pass type I membrane protein (By similarity). Notch 1 intracellular domain: Nucleus (By similarity): P46531
• Total number of polymer chains: 2
• Total formula weight: 54541.45

Authors

Gordon, W.R.,Blacklow, S.C. (deposition date: 2008-10-08, release date: 2008-12-23, Last modification date: 2024-11-06)

Primary citation

Gordon, W.R.,Roy, M.,Vardar-Ulu, D.,Garfinkel, M.,Mansour, M.R.,Aster, J.C.,Blacklow, S.C.
Structure of the Notch1-negative regulatory region: implications for normal activation and pathogenic signaling in T-ALL.
Blood, 113:4381-4390, 2009

PubMed Abstract: Proteolytic resistance of Notch prior to ligand binding depends on the structural integrity of a negative regulatory region (NRR) of the receptor that immediately precedes the transmembrane segment. The NRR includes the 3 Lin12/Notch repeats and the juxtamembrane heterodimerization domain, the region of Notch1 most frequently mutated in T-cell acute lymphoblastic leukemia lymphoma (T-ALL). Here, we report the x-ray structure of the Notch1 NRR in its autoinhibited conformation. A key feature of the Notch1 structure that maintains its closed conformation is a conserved hydrophobic plug that sterically occludes the metalloprotease cleavage site. Crystal packing interactions involving a highly conserved, exposed face on the third Lin12/Notch repeat suggest that this site may normally be engaged in intermolecular or intramolecular protein-protein interactions. The majority of known T-ALL-associated point mutations map to residues in the hydrophobic interior of the Notch1 NRR. A novel mutation (H1545P), which alters a residue at the crystal-packing interface, leads to ligand-independent increases in signaling in reporter gene assays despite only mild destabilization of the NRR, suggesting that it releases the autoinhibitory clamp on the heterodimerization domain imposed by the Lin12/Notch repeats. The Notch1 NRR structure should facilitate a search for antibodies or compounds that stabilize the autoinhibited conformation.

PubMed: 19075186
DOI: 10.1182/blood-2008-08-174748

Experimental method

X-RAY DIFFRACTION (2 Å)