2RR0

Structure of epidermal growth factor-like repeat 12 of mouse Notch-1 receptor

Summary for 2RR0

• Entry DOI: 10.2210/pdb2rr0/pdb
• Related: 1TOZ 2JOA 2RQZ 2RR2
• Descriptor: Neurogenic locus notch homolog protein 1 (1 entity in total)
• Functional Keywords: notch, egf-like domain, activator, ank repeat, cell membrane, developmental protein, differentiation, disulfide bond, glycoprotein, metal-binding, notch signaling pathway, nucleus, phosphoprotein, receptor, transcription, transcription regulation, transmembrane
• Biological source: Mus musculus (MOUSE)
• Cellular location: Cell membrane; Single-pass type I membrane protein. Notch 1 intracellular domain: Nucleus: Q01705
• Total number of polymer chains: 1
• Total formula weight: 4218.70

Authors

Hosoguchi, K.,Shimizu, K.,Fujitani, N.,Nishimura, S. (deposition date: 2010-02-26, release date: 2010-10-13, Last modification date: 2024-10-16)

Primary citation

Hiruma-Shimizu, K.,Hosoguchi, K.,Liu, Y.,Fujitani, N.,Ohta, T.,Hinou, H.,Matsushita, T.,Shimizu, H.,Feizi, T.,Nishimura, S.
Chemical Synthesis, Folding, and Structural Insights into O-Fucosylated Epidermal Growth Factor-like Repeat 12 of Mouse Notch-1 Receptor
J.Am.Chem.Soc., 132:14857-14865, 2010

PubMed Abstract: Notch receptors are cell surface glycoproteins that play key roles in a number of developmental cascades in metazoa. The extracellular domains of Notch-1 receptors are composed of 36 tandem epidermal growth factor (EGF)-like repeats, many of which are modified at highly conserved consensus sites by an unusual form of O-glycan, with O-fucose. The O-fucose residues on certain EGF repeats may be elongated. In mammalian cells this can be a tetrasaccharide, Siaα2,3Galβ1,4GlcNAcβ1,3Fucα1→. This elongation process is initiated by the action of O-fucose-specific β1,3 N-acetylglucosaminyltransferases of the Fringe family. There is evidence that the addition of GlcNAc by Fringe serves as an essential modulator of the interaction of Notch with its ligands and the triggering of activation. Here we describe the efficient synthesis, folding, and structural characterization of EGF repeat 12 (EGF 12) of a mouse Notch-1 receptor bearing different O-fucose glycan chains. We demonstrate that the three disulfide bonds, Cys(456)-Cys(467) (C1-C3), Cys(461)-Cys(476) (C2-C4), and Cys(478)-Cys(487) (C5-C6) were correctly formed in the nonglycosylated as well as the O-fucosylated forms of EGF 12. Three-dimensional structural studies by NMR reveal that the methyl group of fucose is in close contact with ILe(475), Met(477), Pro(478) residues and this stabilizes the conformation of the antiparallel β-sheet of EGF 12. The addition of the GlcNAc residue on O-fucosylated EGF 12 induces a significant conformational change in the adjacent tripeptide sequence, Gln(462)Asn(463)Asp(464), which is a motif involved in the natural, enzymatic O-fucosylation at the conserved site (Cys(461)X(4)Ser/ThrCys(467)).

PubMed: 20883017
DOI: 10.1021/ja105216u

Experimental method

SOLUTION NMR