Structural and functional studies of the EGF20-27 region reveal new features of the human Notch receptor important for optimal activation

DOI: 10.1016/j.str.2024.10.012

Authors: Zhihan Bo (University of Oxford) , Thomas Rowntree (University of Oxford) , Steven Johnson (University of Oxford) , Hilman Nurmahdi (University of Oxford) , Richard J. Suckling (University of Oxford) , Johan Hill (University of Oxford) , Boguslawa Korona (University of Oxford) , Philip C. Weisshuhn (University of Oxford) , Devon Sheppard (University of Oxford) , Yao Meng (University of Oxford) , Shaoyan Liang (University of Oxford) , Edward D. Lowe (University of Oxford) , Susan M. Lea (University of Oxford) , Christina Redfield (University of Oxford) , Penny A. Handford (University of Oxford)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Structure , VOL 284

State: Published (Approved)
Published: November 2024
Diamond Proposal Number(s): 31353 , 7495 , 12346

Abstract: The Notch receptor is activated by the Delta/Serrate/Lag-2 (DSL) family of ligands. The organization of the extracellular signaling complex is unknown, although structures of Notch/ligand complexes comprising the ligand-binding region (LBR), and negative regulatory region (NRR) region, have been solved. Here, we investigate the human Notch-1 epidermal growth factor-like (EGF) 20-27 region, located between the LBR and NRR, and incorporating the Abruptex (Ax) region, associated with distinctive Drosophila phenotypes. Our analyses, using crystallography, NMR and small angle X-ray scattering (SAXS), support a rigid, elongated organization for EGF20-27 with the EGF20-21 linkage showing Ca2+-dependent flexibility. In functional assays, Notch-1 variants containing Ax substitutions result in reduced ligand-dependent trans-activation. When cis-JAG1 was expressed, Notch activity differences between WT and Ca2+-binding Ax variants were less marked than seen in the trans-activation assays alone, consistent with disruption of cis-inhibition. These data indicate the importance of Ca2+-stabilized structure and suggest the balance of cis- and trans-interactions explains the effects of Drosophila Ax mutations.

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: B21-High Throughput SAXS , I03-Macromolecular Crystallography , I04-1-Macromolecular Crystallography (fixed wavelength)

Added On: 04/11/2024 09:43

Documents:
1-s2.0-S0969212624004477-main.pdf

Discipline Tags:

Biochemistry Chemistry Structural biology Life Sciences & Biotech

Technical Tags:

Diffraction Scattering Macromolecular Crystallography (MX) Small Angle X-ray Scattering (SAXS)