Notch-Jagged complex structure implicates a catch bond in tuning ligand sensitivity

Luca, Vincent C.; Kim, Byoung Choul; Ge, Chenghao; Kakuda, Shinako; Wu, Di; Roein-Peikar, Mehdi; Haltiwanger, Robert S.; Zhu, Cheng; Ha, Taekjip; García, K. Christopher

doi:10.1126/science.aaf9739

Cited by 256 publications

(332 citation statements)

References 46 publications

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“…Previous studies have revealed structures of EGF modified by O-fucose and O-glucose glycans (37)(38)(39). We superimposed the current structure of trisaccharide-modified hFA9 with an O-glucose monosaccharide-modified or an O-glucose disaccharide-modified EGF12, and we found that they align well with an r.m.s.d.…”

Section: Structural Analysis Of Hfa9 Egf Repeat Modified With Oglucosmentioning

confidence: 65%

“…Highly ordered O-linked sugar modifications may also be important for molecular recognition because O-linked sugars may work as "surrogate amino acids" in a given protein structure. For example, O-fucose glycans attached to EGF8 and EGF12 of Notch1 interact with the underlying amino acids and form a part of the Notch ligand-binding surface (37)(38)(39)(40). O-Glucose glycans on Notch are not required for Notch-ligand binding (8,16), which is supported by recent co-crystallized structures of Notch1 with ligands (37,38).…”

Section: O-glycans Modulate Egf Protein Stabilitymentioning

confidence: 81%

“…For example, O-fucose glycans attached to EGF8 and EGF12 of Notch1 interact with the underlying amino acids and form a part of the Notch ligand-binding surface (37)(38)(39)(40). O-Glucose glycans on Notch are not required for Notch-ligand binding (8,16), which is supported by recent co-crystallized structures of Notch1 with ligands (37,38). Instead, O-glucose glycans apparently conceal the exposed hydrophobic region of EGF repeats that may otherwise non-specifically bind to unrelated molecules or Notch itself (Fig.…”

Section: O-glycans Modulate Egf Protein Stabilitymentioning

confidence: 99%

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O-Glycosylation modulates the stability of epidermal growth factor-like repeats and thereby regulates Notch trafficking

Takeuchi

Hao

et al. 2017

Journal of Biological Chemistry

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Glycosylation in the endoplasmic reticulum (ER) is closely associated with protein folding and quality control. We recently described a non-canonical ER quality control mechanism for folding of thrombospondin type 1 repeats by protein O-fucosyltransferase 2 (POFUT2). Epidermal growth factor-like (EGF) repeats are also small cysteine-rich protein motifs that can be O-glycosylated by several ER-localized enzymes, including protein O-glucosyltransferase 1 (POGLUT1) and POFUT1. Both POGLUT1 and POFUT1 modify the Notch receptor on multiple EGF repeats and are essential for full Notch function. The fact that POGLUT1 and POFUT1 can distinguish between folded and unfolded EGF repeats raised the possibility that they participate in a quality control pathway for folding of EGF repeats in proteins such as Notch. Here, we demonstrate that cell-surface expression of endogenous Notch1 in HEK293T cells is dependent on the presence of POGLUT1 and POFUT1 in an additive manner. In vitro unfolding assays reveal that addition of O-glucose or O-fucose stabilizes a single EGF repeat and that addition of both O-glucose and O-fucose enhances stability in an additive manner. Finally, we solved the crystal structure of a single EGF repeat covalently modified by a full O-glucose trisaccharide at 2.2 Å resolution. The structure reveals that the glycan fills up a surface groove of the EGF with multiple contacts with the protein, providing a chemical basis for the stabilizing effects of the glycans. Taken together, this work suggests that O-fucose and O-glucose glycans cooperatively stabilize individual EGF repeats through intramolecular interactions, thereby regulating Notch trafficking in cells.

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Section: Structural Analysis Of Hfa9 Egf Repeat Modified With Oglucosmentioning

confidence: 65%

Section: O-glycans Modulate Egf Protein Stabilitymentioning

confidence: 81%

Section: O-glycans Modulate Egf Protein Stabilitymentioning

confidence: 99%

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O-Glycosylation modulates the stability of epidermal growth factor-like repeats and thereby regulates Notch trafficking

Takeuchi

Hao

et al. 2017

Journal of Biological Chemistry

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“…Ligand-specific control of signaling strength may be obtained by posttranslational modifications (68), protein clustering, and localization (69). It was recently demonstrated that different force requirements are needed to activate Notch signaling by different ligands and the authors suggest that this may be a mechanism for Notch-expressing cells to tune their sensitivity to discriminate between different ligands (70). It is possible that a binding between Jagged and vimentin acts as a force-generating mechanism to ensure efficient Jagged-Notch activation.…”

Section: Discussionmentioning

confidence: 99%

Selective regulation of Notch ligands during angiogenesis is mediated by vimentin

Antfolk

Sjöqvist

Cheng

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Notch signaling is a key regulator of angiogenesis, in which sprouting is regulated by an equilibrium between inhibitory Dll4-Notch signaling and promoting Jagged-Notch signaling. Whereas Fringe proteins modify Notch receptors and strengthen their activation by Dll4 ligands, other mechanisms balancing Jagged and Dll4 signaling are yet to be described. The intermediate filament protein vimentin, which has been previously shown to affect vascular integrity and regenerative signaling, is here shown to regulate ligand-specific Notch signaling. Vimentin interacts with Jagged, impedes basal recycling endocytosis of ligands, but is required for efficient receptor ligand transendocytosis and Notch activation upon receptor binding. Analyses of Notch signal activation by using chimeric ligands with swapped intracellular domains (ICDs), demonstrated that the Jagged ICD binds to vimentin and contributes to signaling strength. Vimentin also suppresses expression of Fringe proteins, whereas depletion of vimentin enhances Fringe levels to promote Dll4 signaling. In line with these data, the vasculature in vimentin knockout (VimKO) embryos and placental tissue is underdeveloped with reduced branching. Disrupted angiogenesis in aortic rings from VimKO mice and in endothelial 3D sprouting assays can be rescued by reactivating Notch signaling by recombinant Jagged ligands. Taken together, we reveal a function of vimentin and demonstrate that vimentin regulates Notch ligand signaling activities during angiogenesis.

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“…• The response of cell adhesion complexes to mechanical forces is diverse, [29][30][31]. Phenomenological theories, based on two state models [32,33], and microscopic theory [34,35] …”

Section: Discussionmentioning

confidence: 99%

Forced-rupture of Cell-Adhesion Complexes Reveals abrupt switch between two Brittle States

Toan

Thirumalai

2017

Preprint

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Cell adhesion complexes (CACs), which are activated by ligand binding, play key roles in many cellular functions ranging from cell cycle regulation to mediation of cell extracellular matrix adhesion.Inspired by single molecule pulling experiments using atomic force spectroscopy on leukocyte function-associated antigen-1 (LFA-1), expressed in T-cells, bound to intercellular adhesion molecules (ICAM), we performed constant loading rate (r f ) and constant force (F ) simulations using the Self-Organized Polymer (SOP) model to describe the mechanism of ligand rupture from CACs. The simulations reproduce the major experimental finding on the kinetics of the rupture process, namely, the dependence of the most probable rupture forces (f * s) on ln r f (r f is the loading rate) exhibits two distinct linear regimes. The first, at low r f , has a shallow slope whereas the the slope at high r f is much larger, especially for LFA-1/ICAM-1 complex with the transition between the two occurring over a narrow r f range. Locations of the two transition states (TSs), extracted from the simulations show an abrupt change from a high value at low r f or F to a low value at high r f or F . This unusual behavior in which the CACs switch from one brittle (TS position is a constant over a range of forces) state to another brittle state is not found in forced-rupture in other protein complexes. We explain this novel behavior by constructing the free energy profiles, F (Λ)s, as a function of a collective reaction coordinate (Λ), involving many key charged residues and a critical metal ion 2 peer-reviewed) is the author/funder. All rights reserved. No reuse allowed without permission.The copyright holder for this preprint (which was not . http://dx.doi.org/10.1101/211045 doi: bioRxiv preprint first posted online Oct. 29, 2017; (M g 2+ ). The TS positions in F(Λ), which quantitatively agree with the parameters extracted using the Bell-Evans model, change abruptly at a critical force, demonstrating that it, rather than the molecular extension is a good reaction coordinate. Our combined analyses using simulations performed in both the pulling modes (constant r f and force) reveal a new mechanism for the two loading regimes observed in the rupture kinetics in CACs.

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Notch-Jagged complex structure implicates a catch bond in tuning ligand sensitivity

Cited by 256 publications

References 46 publications

O-Glycosylation modulates the stability of epidermal growth factor-like repeats and thereby regulates Notch trafficking

O-Glycosylation modulates the stability of epidermal growth factor-like repeats and thereby regulates Notch trafficking

Selective regulation of Notch ligands during angiogenesis is mediated by vimentin

Forced-rupture of Cell-Adhesion Complexes Reveals abrupt switch between two Brittle States

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