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Updated: Aug 29, 2025

Cell Aggregation Assays to Evaluate the Binding of the Drosophila Notch with Trans-Ligands and its Inhibition by Cis-Ligands
05:48

Cell Aggregation Assays to Evaluate the Binding of the Drosophila Notch with Trans-Ligands and its Inhibition by Cis-Ligands

Published on: January 2, 2018

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Secretory expression of mammalian NOTCH tandem epidermal growth factor-like repeats based on increased

Ailing Zhang1, Yohei Tsukamoto2, Hideyuki Takeuchi3

  • 1Department of Molecular Biochemistry, Nagoya University Graduate School of Medicine, Nagoya, Japan; Institute for Glyco-core Research (iGCORE), Nagoya University, Nagoya, Japan; Department of Anesthesiology, Nagoya University Graduate School of Medicine, Nagoya, Japan.

Analytical Biochemistry
|September 6, 2022
PubMed

Insights

Enhancing Notch signaling inhibition involves increasing the secretion of Notch EGF repeats. Overexpressing specific glycosyltransferases like EOGT boosts Notch EGF repeat secretion, offering a new strategy for cancer treatment.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Background:

  • The Notch pathway is crucial for intercellular communication and development.
  • Dysregulation of Notch signaling is linked to various diseases, including cancer.
  • Notch EGF repeats can act as decoy molecules to inhibit Notch signaling.

Purpose of the Study:

  • To investigate the effect of glycosyltransferases on the secretion of Notch EGF repeats.
  • To explore strategies for enhancing the production of Notch inhibitory molecules.

Main Methods:

  • Utilized HEK293T cells for overexpression studies.
  • Manipulated the expression of EGF domain-modifying glycosyltransferases, specifically EOGT.
  • Employed label-free glycopeptide quantification to assess O-glycosylation changes.

Main Results:

  • Overexpression of EOGT significantly enhanced the secretion of NOTCH1 EGF repeats.
  • This enhancement was dependent on EOGT enzyme activity.
  • Co-expression with protein O-glucosyltransferase 1 further amplified the effect.

Conclusions:

  • Glycosyltransferase manipulation, particularly EOGT, is a viable strategy to improve Notch EGF repeat secretion.
  • This approach facilitates the efficient generation of Notch inhibitory molecules.
  • The findings offer a novel experimental strategy for cancer treatment development.
Keywords:
EOGTGlycoproteomicsMass spectrometryNOTCHPOGLUT1

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